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Pendergast, Ann Marie

Overview:

The long-term goal of our research is to define the pathways that integrate activation of growth factor, chemokine and adhesion receptors to the regulation of morphogenesis, cell polarity, growth, differentiation, adhesion, and migration during cancer and in response to injury. We have a long-standing research interest on the role of protein tyrosine phosphorylation in tumorigenesis and inflammation. Our early research led to seminal discoveries that defined the critical pathways employed by the Bcr-Abl tyrosine kinase to induce human leukemias. Currently, we are employing novel animal models to investigate the role of tyrosine kinase signaling networks in the regulation of cell polarity, growth, differentiation, adhesion and migration required for tumor progression and inflammatory responses. In particular, we are dissecting the pathways that modulate the crosstalk between multiple cell types during tumor progression and metastasis. Disrupting these “intercellular conversations” is expected to generate new targets for therapeutic intervention. Specifically, we focus on the role of the Abl family of tyrosine kinases, Abl1 and Abl2 (Arg), and associated actin regulatory proteins in diverse cellular processes leading to changes in cell morphology, motility, invasion, adhesion, as well as cell growth and survival. Among the research areas currently being pursued in our laboratory are defining the mechanisms that regulate the cross-talk between cancer cells and associated cells in the tumor microenvironment. We have recently uncovered a previously unknown role for Abl kinases in the regulation of tumor-bone interactions by breast cancer cells and showed that Abl kinases promote breast cancer osteolytic metastasis by activating transcriptional networks dependent on TAZ and STAT5. Moreover, we found that ABL kinases promote metastasis of lung cancer cells harboring EGFR or KRAS mutations. Inactivation of ABL kinases suppresses lung cancer cell metastasis and ABL kinases are required for expression of pro-metastasis genes in lung cancer cells. ABL-mediated activation of the TAZ and b-catenin transcriptional co-activators is required for lung adenocarcinoma metastasis, and ABL kinases activate TAZ- and b-catenin by decreasing their interaction with the b-TrCP ubiquitin ligase leading to increased protein stability. High-level expression of ABL1, ABL2 and a subset of ABL-dependent TAZ- and b-catenin-target genes correlates with shortened survival of lung adenocarcinoma patients. Thus, ABL-specific allosteric inhibitors might be effective to treat metastatic lung cancer with an activated ABL pathway signature. The ultimate goal of our studies is to develop novel therapies for the treatment of metastatic solid tumors by targeting not only cancer cells but also associated stromal cells in the tumor microenvironment.

Positions:

Anthony R. Means Cancer Biology Professor

Pharmacology & Cancer Biology
School of Medicine

Professor of Pharmacology and Cancer Biology

Pharmacology & Cancer Biology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 1986

Ph.D. — University of California at Riverside

Grants:

Medical Scientist Training Program

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1997
End Date
June 30, 2022

Transfusion Medicine and Hematology

Administered By
Medicine, Hematology
AwardedBy
National Institutes of Health
Role
Preceptor
Start Date
July 01, 1975
End Date
June 30, 2021

Novel target for therapy refractory lung tumors

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 08, 2015
End Date
April 30, 2020

Role of ErbB Receptor Signaling in Regulating Normal and Leukemic Stem Cell Fate

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
September 09, 2014
End Date
August 31, 2019

Viral Oncology Training Grant

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Participating Faculty Member
Start Date
July 01, 1980
End Date
June 30, 2019

Regulation of cell permeability by Abl kinases and implications for the treating of acute lung injury

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 2015
End Date
August 31, 2018

Targeting Integrator Kinases in Lung Cancer Metastasis

Administered By
Pharmacology & Cancer Biology
AwardedBy
Free to Breathe
Role
Principal Investigator
Start Date
July 01, 2015
End Date
June 30, 2018

Designing durable apoptosis-targeting therapies for acute myeloid leukemia

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Co-Sponsor
Start Date
May 01, 2016
End Date
April 30, 2018

Role of Abl Kinases in immune cell signaling

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 01, 2004
End Date
June 30, 2017

Kinase Target of Diverse Cell Surface Receptors in Cancer Invasion and Metastasis

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 21, 2011
End Date
April 30, 2017

Cancer Biology Training Grant

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Cancer Institute
Role
Mentor
Start Date
July 01, 1993
End Date
March 31, 2016

Identification of a novel target for treating breast cancer metastasis

Administered By
Pharmacology & Cancer Biology
AwardedBy
American Cancer Society, Inc.
Role
Principal Investigator
Start Date
August 01, 2014
End Date
February 29, 2016

Novel druggable pathway required for lung cancer progression and metastasis

Administered By
Pharmacology & Cancer Biology
AwardedBy
Uniting Against Lung Cancer
Role
Principal Investigator
Start Date
March 01, 2014
End Date
February 28, 2016

Molecular Basis of Signaling by the cAbl Proto-Oncogene

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
March 01, 1996
End Date
February 28, 2013

Intercellular adhesion and morphogenesis: role of actin-regulatory proteins

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 16, 2007
End Date
April 30, 2011

Integrated instrument systems for maintenance and delivery of RNAi libraries

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
February 15, 2008
End Date
February 14, 2009

Signal Amplification by an RTK/Abl Kinase Module

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 15, 2004
End Date
June 30, 2008

Biomarker Studies for Novel Anti-Cancer Agents

Administered By
Medicine, Medical Oncology
AwardedBy
National Institutes of Health
Role
Consultant
Start Date
May 28, 2003
End Date
February 29, 2008

Role of Tyrosine Kinases & Adaptor Proteins in Migration

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 01, 2001
End Date
April 30, 2006

Erythropoietin Receptor Regulation of Erythorpoiesis

Administered By
Medicine, Hematology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1998
End Date
June 30, 2005

Analysis of BCR/ABL Signaling in Human Leukemias

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 1998
End Date
January 31, 2004

Adaptor Proteins And Malignant Transformation

Administered By
Surgery
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1998
End Date
June 30, 2002

ABI-2 A novel C Abl Regulator

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 24, 1997
End Date
September 23, 2000

Bcr/Abl Signaling In Human Leukemias

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 1997
End Date
March 31, 1999

Analysis Of Bcr/Abl Signaling In Human Leukemias

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
June 15, 1994
End Date
March 31, 1999

Molecular Basis Of Signaling By The Cab1 Proto-Oncogene

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 15, 1996
End Date
February 28, 1999

Molecular Basis Of Signaling By The Cabl Proto-Oncogene

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 15, 1996
End Date
February 28, 1999
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Awards:

AAAS Fellows. American Association for the Advancement of Science, The.

Type
National
Awarded By
American Association for the Advancement of Science, The
Date
January 01, 2010

Publications:

Inactivation of ABL kinases suppresses non-small cell lung cancer metastasis.

Current therapies to treat non-small cell lung carcinoma (NSCLC) have proven ineffective owing to transient, variable, and incomplete responses. Here we show that ABL kinases, ABL1 and ABL2, promote metastasis of lung cancer cells harboring EGFR or KRAS mutations. Inactivation of ABL kinases suppresses NSCLC metastasis to brain and bone, and other organs. ABL kinases are required for expression of prometastasis genes. Notably, ABL1 and ABL2 depletion impairs extravasation of lung adenocarcinoma cells into the lung parenchyma. We found that ABL-mediated activation of the TAZ and β-catenin transcriptional coactivators is required for NSCLC metastasis. ABL kinases activate TAZ and β-catenin by decreasing their interaction with the β-TrCP ubiquitin ligase, leading to increased protein stability. High-level expression of ABL1, ABL2, and a subset of ABL-dependent TAZ- and β-catenin-target genes correlates with shortened survival of lung adenocarcinoma patients. Thus, ABL-specific allosteric inhibitors might be effective to treat metastatic lung cancer with an activated ABL pathway signature.

Authors
Gu, JJ; Rouse, C; Xu, X; Wang, J; Onaitis, MW; Pendergast, AM
MLA Citation
Gu, JJ, Rouse, C, Xu, X, Wang, J, Onaitis, MW, and Pendergast, AM. "Inactivation of ABL kinases suppresses non-small cell lung cancer metastasis." JCI insight 1.21 (December 22, 2016): e89647-.
PMID
28018973
Source
epmc
Published In
JCI insight
Volume
1
Issue
21
Publish Date
2016
Start Page
e89647
DOI
10.1172/jci.insight.89647

Reciprocal stabilization of ABL and TAZ regulates osteoblastogenesis through transcription factor RUNX2.

Cellular identity in metazoan organisms is frequently established through lineage-specifying transcription factors, which control their own expression through transcriptional positive feedback, while antagonizing the developmental networks of competing lineages. Here, we have uncovered a distinct positive feedback loop that arises from the reciprocal stabilization of the tyrosine kinase ABL and the transcriptional coactivator TAZ. Moreover, we determined that this loop is required for osteoblast differentiation and embryonic skeletal formation. ABL potentiated the assembly and activation of the RUNX2-TAZ master transcription factor complex that is required for osteoblastogenesis, while antagonizing PPARγ-mediated adipogenesis. ABL also enhanced TAZ nuclear localization and the formation of the TAZ-TEAD complex that is required for osteoblast expansion. Last, we have provided genetic data showing that regulation of the ABL-TAZ amplification loop lies downstream of the adaptor protein 3BP2, which is mutated in the craniofacial dysmorphia syndrome cherubism. Our study demonstrates an interplay between ABL and TAZ that controls the mesenchymal maturation program toward the osteoblast lineage and is mechanistically distinct from the established model of lineage-specific maturation.

Authors
Matsumoto, Y; La Rose, J; Kent, OA; Wagner, MJ; Narimatsu, M; Levy, AD; Omar, MH; Tong, J; Krieger, JR; Riggs, E; Storozhuk, Y; Pasquale, J; Ventura, M; Yeganeh, B; Post, M; Moran, MF; Grynpas, MD; Wrana, JL; Superti-Furga, G; Koleske, AJ; Pendergast, AM; Rottapel, R
MLA Citation
Matsumoto, Y, La Rose, J, Kent, OA, Wagner, MJ, Narimatsu, M, Levy, AD, Omar, MH, Tong, J, Krieger, JR, Riggs, E, Storozhuk, Y, Pasquale, J, Ventura, M, Yeganeh, B, Post, M, Moran, MF, Grynpas, MD, Wrana, JL, Superti-Furga, G, Koleske, AJ, Pendergast, AM, and Rottapel, R. "Reciprocal stabilization of ABL and TAZ regulates osteoblastogenesis through transcription factor RUNX2." The Journal of clinical investigation 126.12 (December 2016): 4482-4496.
PMID
27797343
Source
epmc
Published In
Journal of Clinical Investigation
Volume
126
Issue
12
Publish Date
2016
Start Page
4482
End Page
4496
DOI
10.1172/jci87802

ABL kinases promote breast cancer osteolytic metastasis by modulating tumor-bone interactions through TAZ and STAT5 signaling.

Bone metastases occur in up to 70% of advanced breast cancer. For most patients with breast cancer, bone metastases are predominantly osteolytic. Interactions between tumor cells and stromal cells in the bone microenvironment drive osteolytic bone metastasis, a process that requires the activation of osteoclasts, cells that break down bone. We report that ABL kinases promoted metastasis of breast cancer cells to bone by regulating the crosstalk between tumor cells and the bone microenvironment. ABL kinases protected tumor cells from apoptosis induced by TRAIL (TNF-related apoptosis-inducing ligand), activated the transcription factor STAT5, and promoted osteolysis through the STAT5-dependent expression of genes encoding the osteoclast-activating factors interleukin-6 (IL-6) and matrix metalloproteinase 1 (MMP1). Furthermore, in breast cancer cells, ABL kinases increased the abundance of the Hippo pathway mediator TAZ and the expression of TAZ-dependent target genes that promote bone metastasis. Knockdown of ABL kinases or treatment with ABL-specific allosteric inhibitor impaired osteolytic metastasis of breast cancer cells in mice. These findings revealed a role for ABL kinases in regulating tumor-bone interactions and provide a rationale for using ABL-specific inhibitors to limit breast cancer metastasis to bone.

Authors
Wang, J; Rouse, C; Jasper, JS; Pendergast, AM
MLA Citation
Wang, J, Rouse, C, Jasper, JS, and Pendergast, AM. "ABL kinases promote breast cancer osteolytic metastasis by modulating tumor-bone interactions through TAZ and STAT5 signaling." Science signaling 9.413 (February 2, 2016): ra12-.
PMID
26838548
Source
epmc
Published In
Science Signaling
Volume
9
Issue
413
Publish Date
2016
Start Page
ra12
DOI
10.1126/scisignal.aad3210

Multifunctional Abl kinases in health and disease.

The Abelson tyrosine kinases were initially identified as drivers of leukemia in mice and humans. The Abl family kinases Abl1 and Abl2 regulate diverse cellular processes during development and normal homeostasis, and their functions are subverted during inflammation, cancer and other pathologies. Abl kinases can be activated by multiple stimuli leading to cytoskeletal reorganization required for cell morphogenesis, motility, adhesion and polarity. Depending on the cellular context, Abl kinases regulate cell survival and proliferation. Emerging data support important roles for Abl kinases in pathologies linked to inflammation. Among these are neurodegenerative diseases and inflammatory pathologies. Unexpectedly, Abl kinases have also been identified as important players in mammalian host cells during microbial pathogenesis. Thus, the use of Abl kinase inhibitors might prove to be effective in the treatment of pathologies beyond leukemia and solid tumors. In this Cell Science at a Glance article and in the accompanying poster, we highlight the emerging roles of Abl kinases in the regulation of cellular processes in normal cells and diverse pathologies ranging from cancer to microbial pathogenesis.

Authors
Khatri, A; Wang, J; Pendergast, AM
MLA Citation
Khatri, A, Wang, J, and Pendergast, AM. "Multifunctional Abl kinases in health and disease." Journal of cell science 129.1 (January 2016): 9-16. (Review)
PMID
26729027
Source
epmc
Published In
Journal of cell science
Volume
129
Issue
1
Publish Date
2016
Start Page
9
End Page
16
DOI
10.1242/jcs.175521

The Emerging Role of ABL Kinases in Solid Tumors.

The Abelson (ABL) tyrosine kinases were identified as drivers of leukemia in mice and humans. Emerging data has shown a role for the ABL family kinases, ABL1 and ABL2, in the progression of several solid tumors. This review will focus on recent reports of the involvement of the ABL kinases in tumor progression using mouse models as well as recent data generated from genomic and proteomic studies linking enhanced expression and hyper-activation of the ABL kinases to some human cancers. Preclinical studies on small molecule inhibitors of the ABL kinases suggest that their use may have beneficial effects for the treatment of selected solid tumors.

Authors
Wang, J; Pendergast, AM
MLA Citation
Wang, J, and Pendergast, AM. "The Emerging Role of ABL Kinases in Solid Tumors." Trends in cancer 1.2 (October 2015): 110-123.
PMID
26645050
Source
epmc
Published In
Trends in cancer
Volume
1
Issue
2
Publish Date
2015
Start Page
110
End Page
123

ShcA regulates thymocyte proliferation through specific transcription factors and a c-Abl-dependent signaling axis.

Signaling via the pre-T-cell receptor (pre-TCR), along with associated signals from Notch and chemokine receptors, regulates the β-selection checkpoint that operates on CD4(-) CD8(-) doubly negative (DN) thymocytes. Since many hematopoietic malignancies arise at the immature developmental stages of lymphocytes, understanding the signal integration and how specific signaling molecules and distal transcription factors regulate cellular outcomes is of importance. Here, a series of molecular and genetic approaches revealed that the ShcA adapter protein critically influences proliferation and differentiation during β-selection. We found that ShcA functions downstream of the pre-TCR and p56(Lck) and show that ShcA is important for extracellular signal-regulated kinase (ERK)-dependent upregulation of transcription factors early growth factor 1 (Egr1) and Egr3 in immature thymocytes and, in turn, of the expression and function of the Id3 and E2A helix-loop-helix (HLH) proteins. ShcA also contributes to pre-TCR-mediated induction of c-Myc and additional cell cycle regulators. Moreover, using an unbiased Saccharomyces cerevisiae (yeast) screen, we identified c-Abl as a binding partner of phosphorylated ShcA and demonstrated the relevance of the ShcA-c-Abl interaction in immature thymocytes. Collectively, these data identify multiple modes by which ShcA can fine-tune the development of early thymocytes, including a previously unappreciated ShcA-c-Abl axis that regulates thymocyte proliferation.

Authors
Trampont, PC; Zhang, L; Giles, AJ; Walk, SF; Gu, JJ; Pendergast, AM; Ravichandran, KS
MLA Citation
Trampont, PC, Zhang, L, Giles, AJ, Walk, SF, Gu, JJ, Pendergast, AM, and Ravichandran, KS. "ShcA regulates thymocyte proliferation through specific transcription factors and a c-Abl-dependent signaling axis." Molecular and cellular biology 35.8 (April 2015): 1462-1476.
PMID
25691660
Source
epmc
Published In
Molecular and Cellular Biology
Volume
35
Issue
8
Publish Date
2015
Start Page
1462
End Page
1476
DOI
10.1128/mcb.01084-14

Abl Kinases Regulate HGF/Met Signaling Required for Epithelial Cell Scattering, Tubulogenesis and Motility.

Tight regulation of receptor tyrosine kinases (RTKs) is crucial for normal development and homeostasis. Dysregulation of RTKs signaling is associated with diverse pathological conditions including cancer. The Met RTK is the receptor for hepatocyte growth factor (HGF) and is dysregulated in numerous human tumors. Here we show that Abl family of non-receptor tyrosine kinases, comprised of Abl (ABL1) and Arg (ABL2), are activated downstream of the Met receptor, and that inhibition of Abl kinases dramatically suppresses HGF-induced cell scattering and tubulogenesis. We uncover a critical role for Abl kinases in the regulation of HGF/Met-dependent RhoA activation and RhoA-mediated actomyosin contractility and actin cytoskeleton remodeling in epithelial cells. Moreover, treatment of breast cancer cells with Abl inhibitors markedly decreases Met-driven cell migration and invasion. Notably, expression of a transforming mutant of the Met receptor in the mouse mammary epithelium results in hyper-activation of both Abl and Arg kinases. Together these data demonstrate that Abl kinases link Met activation to Rho signaling and Abl kinases are required for Met-dependent cell scattering, tubulogenesis, migration, and invasion. Thus, inhibition of Abl kinases might be exploited for the treatment of cancers driven by hyperactivation of HGF/Met signaling.

Authors
Li, R; Knight, JF; Park, M; Pendergast, AM
MLA Citation
Li, R, Knight, JF, Park, M, and Pendergast, AM. "Abl Kinases Regulate HGF/Met Signaling Required for Epithelial Cell Scattering, Tubulogenesis and Motility." PloS one 10.5 (January 2015): e0124960-.
PMID
25946048
Source
epmc
Published In
PloS one
Volume
10
Issue
5
Publish Date
2015
Start Page
e0124960
DOI
10.1371/journal.pone.0124960

Targeting ABL1-mediated oxidative stress adaptation in fumarate hydratase-deficient cancer.

Patients with germline fumarate hydratase (FH) mutation are predisposed to develop aggressive kidney cancer with few treatment options and poor therapeutic outcomes. Activity of the proto-oncogene ABL1 is upregulated in FH-deficient kidney tumors and drives a metabolic and survival signaling network necessary to cope with impaired mitochondrial function and abnormal accumulation of intracellular fumarate. Excess fumarate indirectly stimulates ABL1 activity, while restoration of wild-type FH abrogates both ABL1 activation and the cytotoxicity caused by ABL1 inhibition or knockdown. ABL1 upregulates aerobic glycolysis via the mTOR/HIF1α pathway and neutralizes fumarate-induced proteotoxic stress by promoting nuclear localization of the antioxidant response transcription factor NRF2. Our findings identify ABL1 as a pharmacologically tractable therapeutic target in glycolytically dependent, oxidatively stressed tumors.

Authors
Sourbier, C; Ricketts, CJ; Matsumoto, S; Crooks, DR; Liao, P-J; Mannes, PZ; Yang, Y; Wei, M-H; Srivastava, G; Ghosh, S; Chen, V; Vocke, CD; Merino, M; Srinivasan, R; Krishna, MC; Mitchell, JB; Pendergast, AM; Rouault, TA; Neckers, L; Linehan, WM
MLA Citation
Sourbier, C, Ricketts, CJ, Matsumoto, S, Crooks, DR, Liao, P-J, Mannes, PZ, Yang, Y, Wei, M-H, Srivastava, G, Ghosh, S, Chen, V, Vocke, CD, Merino, M, Srinivasan, R, Krishna, MC, Mitchell, JB, Pendergast, AM, Rouault, TA, Neckers, L, and Linehan, WM. "Targeting ABL1-mediated oxidative stress adaptation in fumarate hydratase-deficient cancer." Cancer cell 26.6 (December 2014): 840-850.
PMID
25490448
Source
epmc
Published In
Cancer Cell
Volume
26
Issue
6
Publish Date
2014
Start Page
840
End Page
850
DOI
10.1016/j.ccell.2014.10.005

Tie2 (to) Abl: Signaling to endothelial cell survival.

Authors
Chislock, EM; Pendergast, AM
MLA Citation
Chislock, EM, and Pendergast, AM. "Tie2 (to) Abl: Signaling to endothelial cell survival." Cell Cycle 12.24 (December 15, 2013): 3709-3710.
PMID
24145226
Source
pubmed
Published In
Cell Cycle
Volume
12
Issue
24
Publish Date
2013
Start Page
3709
End Page
3710
DOI
10.4161/cc.26877

Role of ABL family kinases in cancer: from leukaemia to solid tumours.

The Abelson (ABL) family of nonreceptor tyrosine kinases, ABL1 and ABL2, transduces diverse extracellular signals to protein networks that control proliferation, survival, migration and invasion. ABL1 was first identified as an oncogene required for the development of leukaemias initiated by retroviruses or chromosome translocations. The demonstration that small-molecule ABL kinase inhibitors could effectively treat chronic myeloid leukaemia opened the door to the era of targeted cancer therapies. Recent reports have uncovered roles for ABL kinases in solid tumours. Enhanced ABL expression and activation in some solid tumours, together with altered cell polarity, invasion or growth induced by activated ABL kinases, suggest that drugs targeting these kinases may be useful for treating selected solid tumours.

Authors
Greuber, EK; Smith-Pearson, P; Wang, J; Pendergast, AM
MLA Citation
Greuber, EK, Smith-Pearson, P, Wang, J, and Pendergast, AM. "Role of ABL family kinases in cancer: from leukaemia to solid tumours." Nat Rev Cancer 13.8 (August 2013): 559-571. (Review)
PMID
23842646
Source
pubmed
Published In
Nature Reviews Cancer
Volume
13
Issue
8
Publish Date
2013
Start Page
559
End Page
571
DOI
10.1038/nrc3563

Abl kinases are required for vascular function, Tie2 expression, and angiopoietin-1-mediated survival.

Endothelial dysfunction is associated with diverse cardiovascular pathologies. Here, we show a previously unappreciated role for the Abelson (Abl) family kinases (Abl and Arg) in endothelial function and the regulation of angiogenic factor pathways important for vascular homeostasis. Endothelial Abl deletion in Arg-null mice led to late-stage embryonic and perinatal lethality, with mutant mice displaying focal loss of vasculature and tissue necrosis. Loss of Abl kinases led to increased endothelial cell apoptosis both in vitro and in vivo, contributing to vascular dysfunction, infarction, and tissue damage. Mechanistically, we identify a unique dual role for Abl kinases in the regulation of angiopoietin/Tie2 protein kinase signaling. Endothelial Abl kinases modulate Tie2 expression and angiopoietin-1-mediated endothelial cell survival. These findings reveal a critical requirement for the Abl kinases in vascular development and function, which may have important implications for the clinical use of Abl kinase inhibitors.

Authors
Chislock, EM; Ring, C; Pendergast, AM
MLA Citation
Chislock, EM, Ring, C, and Pendergast, AM. "Abl kinases are required for vascular function, Tie2 expression, and angiopoietin-1-mediated survival." Proc Natl Acad Sci U S A 110.30 (July 23, 2013): 12432-12437.
PMID
23840065
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
110
Issue
30
Publish Date
2013
Start Page
12432
End Page
12437
DOI
10.1073/pnas.1304188110

Abl family kinases regulate endothelial barrier function in vitro and in mice.

The maintenance of endothelial barrier function is essential for normal physiology, and increased vascular permeability is a feature of a wide variety of pathological conditions, leading to complications including edema and tissue damage. Use of the pharmacological inhibitor imatinib, which targets the Abl family of non-receptor tyrosine kinases (Abl and Arg), as well as other tyrosine kinases including the platelet-derived growth factor receptor (PDGFR), Kit, colony stimulating factor 1 receptor (CSF1R), and discoidin domain receptors, has shown protective effects in animal models of inflammation, sepsis, and other pathologies characterized by enhanced vascular permeability. However, the imatinib targets involved in modulation of vascular permeability have not been well-characterized, as imatinib inhibits multiple tyrosine kinases not only in endothelial cells and pericytes but also immune cells important for disorders associated with pathological inflammation and abnormal vascular permeability. In this work we employ endothelial Abl knockout mice to show for the first time a direct role for Abl in the regulation of vascular permeability in vivo. Using both Abl/Arg-specific pharmacological inhibition and endothelial Abl knockout mice, we demonstrate a requirement for Abl kinase activity in the induction of endothelial permeability by vascular endothelial growth factor both in vitro and in vivo. Notably, Abl kinase inhibition also impaired endothelial permeability in response to the inflammatory mediators thrombin and histamine. Mechanistically, we show that loss of Abl kinase activity was accompanied by activation of the barrier-stabilizing GTPases Rac1 and Rap1, as well as inhibition of agonist-induced Ca(2+) mobilization and generation of acto-myosin contractility. In all, these findings suggest that pharmacological targeting of the Abl kinases may be capable of inhibiting endothelial permeability induced by a broad range of agonists and that use of Abl kinase inhibitors may have potential for the treatment of disorders involving pathological vascular leakage.

Authors
Chislock, EM; Pendergast, AM
MLA Citation
Chislock, EM, and Pendergast, AM. "Abl family kinases regulate endothelial barrier function in vitro and in mice. (Published online)" PLoS One 8.12 (2013): e85231-.
PMID
24367707
Source
pubmed
Published In
PloS one
Volume
8
Issue
12
Publish Date
2013
Start Page
e85231
DOI
10.1371/journal.pone.0085231

Abl family kinases regulate FcγR-mediated phagocytosis in murine macrophages.

Phagocytosis of Ab-coated pathogens is mediated through FcγRs, which activate intracellular signaling pathways to drive actin cytoskeletal rearrangements. Abl and Arg define a family of nonreceptor tyrosine kinases that regulate actin-dependent processes in a variety of cell types, including those important in the adaptive immune response. Using pharmacological inhibition as well as dominant negative and knockout approaches, we demonstrate a role for the Abl family kinases in phagocytosis by macrophages and define a mechanism whereby Abl kinases regulate this process. Bone marrow-derived macrophages from mice lacking Abl and Arg kinases exhibit inefficient phagocytosis of sheep erythrocytes and zymosan particles. Treatment with the Abl kinase inhibitors imatinib and GNF-2 or overexpression of kinase-inactive forms of the Abl family kinases also impairs particle internalization in murine macrophages, indicating Abl kinase activity is required for efficient phagocytosis. Further, Arg kinase is present at the phagocytic cup, and Abl family kinases are activated by FcγR engagement. The regulation of phagocytosis by Abl family kinases is mediated in part by the spleen tyrosine kinase (Syk). Loss of Abl and Arg expression or treatment with Abl inhibitors reduced Syk phosphorylation in response to FcγR ligation. The link between Abl family kinases and Syk may be direct, as purified Arg kinase phosphorylates Syk in vitro. Further, overexpression of membrane-targeted Syk in cells treated with Abl kinase inhibitors partially rescues the impairment in phagocytosis. Together, these findings reveal that Abl family kinases control the efficiency of phagocytosis in part through the regulation of Syk function.

Authors
Greuber, EK; Pendergast, AM
MLA Citation
Greuber, EK, and Pendergast, AM. "Abl family kinases regulate FcγR-mediated phagocytosis in murine macrophages." J Immunol 189.11 (December 1, 2012): 5382-5392.
PMID
23100514
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
189
Issue
11
Publish Date
2012
Start Page
5382
End Page
5392
DOI
10.4049/jimmunol.1200974

Abl family kinases modulate T cell-mediated inflammation and chemokine-induced migration through the adaptor HEF1 and the GTPase Rap1.

Chemokine signaling is critical for T cell function during homeostasis and inflammation and directs T cell polarity and migration through the activation of specific intracellular pathways. Here, we uncovered a previously uncharacterized role for the Abl family tyrosine kinases Abl and Arg in the regulation of T cell-dependent inflammatory responses and showed that the Abl family kinases were required for chemokine-induced T cell polarization and migration. Our data demonstrated that Abl and Arg were activated downstream of chemokine receptors and mediated the chemokine-induced tyrosine phosphorylation of human enhancer of filamentation 1 (HEF1), an adaptor protein that is required for the activity of the guanosine triphosphatase Rap1, which mediates cell adhesion and migration. Phosphorylation of HEF1 by Abl family kinases and activation of Rap1 were required for chemokine-induced T cell migration. Mouse T cells that lacked Abl and Arg exhibited defective homing to lymph nodes and impaired migration to sites of inflammation. These findings suggest that Abl family kinases are potential therapeutic targets for the treatment of T cell-dependent immune disorders that are characterized by chemokine-mediated inflammation.

Authors
Gu, JJ; Lavau, CP; Pugacheva, E; Soderblom, EJ; Moseley, MA; Pendergast, AM
MLA Citation
Gu, JJ, Lavau, CP, Pugacheva, E, Soderblom, EJ, Moseley, MA, and Pendergast, AM. "Abl family kinases modulate T cell-mediated inflammation and chemokine-induced migration through the adaptor HEF1 and the GTPase Rap1. (Published online)" Sci Signal 5.233 (July 17, 2012): ra51-.
PMID
22810897
Source
pubmed
Published In
Science Signaling
Volume
5
Issue
233
Publish Date
2012
Start Page
ra51
DOI
10.1126/scisignal.2002632

ShcA and c-Abl provide a new signaling axis regulating beta-selection

Authors
Trampont, P; Zhang, L; Giles, A; Walk, S; Pendergast, AM; Ravichandran, K
MLA Citation
Trampont, P, Zhang, L, Giles, A, Walk, S, Pendergast, AM, and Ravichandran, K. "ShcA and c-Abl provide a new signaling axis regulating beta-selection." May 1, 2012.
Source
wos-lite
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
188
Publish Date
2012

Arg kinase regulates epithelial cell polarity by targeting β1-integrin and small GTPase pathways.

BACKGROUND: Establishment and maintenance of epithelial cell polarity is regulated in part by signaling from adhesion receptors. Loss of cell polarity is associated with multiple pathologies including the initiation and progression of various cancers. The β1-integrin adhesion receptor plays a role in the regulation of cell polarity; however, the identity of the signaling pathways that modulate β1-integrin function and connect it to the regulation of polarity pathways remains largely unknown. RESULTS: The present work identifies a role for Arg, a member of the Abl family nonreceptor tyrosine kinases, in the regulation of adhesive signals and epithelial cell polarity. In a three-dimensional cell culture model, activation of Arg kinase leads to a striking inversion of apical-basal polarity. In contrast, loss of Arg function impairs the establishment of a polarized epithelial cyst structure. Activated Arg kinase disrupts β1-integrin signaling and localization and impairs Rac1-mediated laminin assembly. Disruption of β1-integrin function by active Arg results in altered distribution of selected polarity complex components mediated in part by Rap1 GTPase signaling. Whereas polarity inversion is partially rescued by a constitutively active Rap1, Rac1-dependent laminin assembly is not, indicating that Rap1 and Rac1 signal independently during epithelial polarity. CONCLUSIONS: These findings suggest that modulation of Arg kinase function may contribute not only to normal epithelial polarity regulation but also may promote pathologies associated with loss of cell polarity.

Authors
Li, R; Pendergast, AM
MLA Citation
Li, R, and Pendergast, AM. "Arg kinase regulates epithelial cell polarity by targeting β1-integrin and small GTPase pathways." Curr Biol 21.18 (September 27, 2011): 1534-1542.
PMID
21906945
Source
pubmed
Published In
Current Biology
Volume
21
Issue
18
Publish Date
2011
Start Page
1534
End Page
1542
DOI
10.1016/j.cub.2011.08.023

Abl-interactor-1 (Abi1) has a role in cardiovascular and placental development and is a binding partner of the alpha4 integrin.

Dynamic signals linking the actin cytoskeleton and cell adhesion receptors are essential for morphogenesis during development and normal tissue homeostasis. Abi1 is a central regulator of actin polymerization through interactions with multiple protein complexes. However, the in vivo role of Abi1 remains to be defined. The α4 integrin adhesion receptor is associated with enhanced protrusive activity and regulation of directional cell migration. Among integrin subunits, α4 exhibits unique properties in that it predominantly accumulates at the leading edge of migrating cells; however, the pathways that link the actin-regulatory machinery to α4 at the leading edge have remained elusive. We generated Abi1 KO mice and found that loss of Abi1 phenocopies KO of α4. Mice lacking Abi1 or α4 exhibit midgestational lethality with abnormalities in placental and cardiovascular development. Notably, purified Abi1 protein binds directly to the α4 cytoplasmic tail and endogenous Abi1 colocalizes with phosphorylated α4 at the leading edge of spreading cells. Moreover, Abi1-deficient cells expressing α4 have impaired cell spreading, which is rescued by WT Abi1 but not an Abi1 mutant lacking the α4-binding site. These data reveal a direct link between the α4 integrin and actin polymerization and uncover a role for Abi1 in the regulation of morphogenesis in vivo. The Abi1-α4 interaction establishes a mechanistic paradigm for signaling between adhesion events and enhanced actin polymerization at the earliest stages of protrusion.

Authors
Ring, C; Ginsberg, MH; Haling, J; Pendergast, AM
MLA Citation
Ring, C, Ginsberg, MH, Haling, J, and Pendergast, AM. "Abl-interactor-1 (Abi1) has a role in cardiovascular and placental development and is a binding partner of the alpha4 integrin." Proc Natl Acad Sci U S A 108.1 (January 4, 2011): 149-154.
PMID
21173240
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
108
Issue
1
Publish Date
2011
Start Page
149
End Page
154
DOI
10.1073/pnas.1012316108

Abl kinases are required for invadopodia formation and chemokine-induced invasion.

The Abl tyrosine kinases, Abl and Arg, play a role in the regulation of the actin cytoskeleton by modulating cell-cell adhesion and cell motility. Deregulation of both the actin cytoskeleton and Abl kinases have been implicated in cancers. Abl kinase activity is elevated in a number of metastatic cancers and these kinases are activated downstream of several oncogenic growth factor receptor signaling pathways. However, the role of Abl kinases in regulation of the actin cytoskeleton during tumor progression and invasion remains elusive. Here we identify the Abl kinases as essential regulators of invadopodia assembly and function. We show that Abl kinases are activated downstream of the chemokine receptor, CXCR4, and are required for cancer cell invasion and matrix degradation induced by SDF1α, serum growth factors, and activated Src kinase. Moreover, Abl kinases are readily detected at invadopodia assembly sites and their inhibition prevents the assembly of actin and cortactin into organized invadopodia structures. We show that active Abl kinases form complexes with membrane type-1 matrix metalloproteinase (MT1-MMP), a critical invadopodia component required for matrix degradation. Further, loss of Abl kinase signaling induces internalization of MT1-MMP from the cell surface, promotes its accumulation in the perinuclear compartment and inhibits MT1-MMP tyrosine phosphorylation. Our findings reveal that Abl kinase signaling plays a critical role in invadopodia formation and function, and have far-reaching implications for the treatment of metastatic carcinomas.

Authors
Smith-Pearson, PS; Greuber, EK; Yogalingam, G; Pendergast, AM
MLA Citation
Smith-Pearson, PS, Greuber, EK, Yogalingam, G, and Pendergast, AM. "Abl kinases are required for invadopodia formation and chemokine-induced invasion." J Biol Chem 285.51 (December 17, 2010): 40201-40211.
PMID
20937825
Source
pubmed
Published In
The Journal of biological chemistry
Volume
285
Issue
51
Publish Date
2010
Start Page
40201
End Page
40211
DOI
10.1074/jbc.M110.147330

Regulation of cell-cell adhesion by Abi/Diaphanous complexes.

Actin polymerization provides the driving force for the formation of cell-cell junctions and is mediated by two types of actin nucleators, Arp2/3 and formins. Proteins required for coordinately linking cadherin-mediated adhesion to Arp2/3-dependent versus formin-dependent nucleation have yet to be defined. Here we show a role for Abi, the Abi-binding partner Nap1, and the Nap1-binding protein Sra1 in the regulation of cadherin-dependent adhesion. We found that Abi, which is known to interact with Wave, leading to activation of the Arp2/3 complex, is also capable of interacting with the Diaphanous (Dia)-related formins in the absence of Wave. Knockdown of Abi, Nap1, Sra1, or Dia markedly inhibited cell-cell junctions, whereas knockdown of Wave or Arp2/3 produced mild and transient phenotypes. Dia and Abi colocalized with beta-catenin at cell-cell junctions. Further, Dia and Wave bound to overlapping sites on Abi1, and Wave competed with Dia for Abi1 binding. Notably, an active Dia1 C-terminal fragment that localizes to cell-cell junctions rescued the abnormal junctions induced by depletion of Abi or Nap1 in epithelial cells. These findings uncover a novel link between cadherin-mediated adhesion and the regulation of actin dynamics through the requirement for an Abi/Dia complex for the formation and stability of cell-cell junctions.

Authors
Ryu, JR; Echarri, A; Li, R; Pendergast, AM
MLA Citation
Ryu, JR, Echarri, A, Li, R, and Pendergast, AM. "Regulation of cell-cell adhesion by Abi/Diaphanous complexes." Mol Cell Biol 29.7 (April 2009): 1735-1748.
PMID
19158278
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
29
Issue
7
Publish Date
2009
Start Page
1735
End Page
1748
DOI
10.1128/MCB.01483-08

Abl tyrosine kinases in T-cell signaling.

Stimulation of the T-cell antigen receptor (TCR) leads to the activation of signaling pathways that are essential for T-cell development and the response of mature T cells to antigens. The TCR has no intrinsic catalytic activity, but TCR engagement results in tyrosine phosphorylation of downstream targets by non-receptor tyrosine kinases. Three families of tyrosine kinases have long been recognized to play critical roles in TCR-dependent signaling. They are the Src, zeta-associated protein of 70 kDa, and Tec families of kinases. More recently, the Abelson (Abl) tyrosine kinases have been shown to be activated by TCR engagement and to be required for maximal TCR signaling. Using T-cell conditional knockout mice deficient for Abl family kinases, Abl (Abl1) and Abl-related gene (Arg) (Abl2), it was recently shown that loss of Abl kinases results in defective T-cell development and a partial block in the transition to the CD4(+)CD8(+) stage. Abl/Arg double null T cells exhibit impaired TCR-induced signaling, proliferation, and cytokine production. Moreover, conditional knockout mice lacking Abl and Arg in T cells exhibit impaired CD8(+) T-cell expansion in vivo upon Listeria monocytogenes infection. Thus, Abl kinase signaling is required for both T-cell development and mature T-cell function.

Authors
Gu, JJ; Ryu, JR; Pendergast, AM
MLA Citation
Gu, JJ, Ryu, JR, and Pendergast, AM. "Abl tyrosine kinases in T-cell signaling." Immunol Rev 228.1 (March 2009): 170-183. (Review)
PMID
19290927
Source
pubmed
Published In
Immunological Reviews
Volume
228
Issue
1
Publish Date
2009
Start Page
170
End Page
183
DOI
10.1111/j.1600-065X.2008.00751.x

Abl kinases regulate autophagy by promoting the trafficking and function of lysosomal components.

Autophagy is a lysosome-dependent degradative pathway that regulates the turnover of intracellular organelles, parasites, and long-lived proteins. Deregulation of autophagy results in a variety of pathological conditions, but little is known regarding the mechanisms that link normal cellular and pathological signals to the regulation of distinct stages in the autophagy pathway. Here we uncover a novel role for the Abl family kinases in the regulation of the late stages of autophagy. Inhibition, depletion, or knockout of the Abl family kinases, Abl and Arg, resulted in a dramatic reduction in the intracellular activities of the lysosomal glycosidases alpha-galactosidase, alpha-mannosidase and neuraminidase. Inhibition of Abl kinases also reduced the processing of the precursor forms of cathepsin D and cathepsin L to their mature, lysosomal forms, which coincided with the impaired turnover of long-lived cytosolic proteins and accumulation of autophagosomes. Furthermore, defective lysosomal degradation of long-lived proteins in the absence of Abl kinase signaling was accompanied by a perinuclear redistribution of lysosomes and increased glycosylation and stability of lysosome-associated membrane proteins, which are known to be substrates for lysosomal enzymes and play a role in regulating lysosome mobility. Our findings reveal a role for Abl kinases in the regulation of late-stage autophagy and have important implications for therapies that employ pharmacological inhibitors of the Abl kinases.

Authors
Yogalingam, G; Pendergast, AM
MLA Citation
Yogalingam, G, and Pendergast, AM. "Abl kinases regulate autophagy by promoting the trafficking and function of lysosomal components." J Biol Chem 283.51 (December 19, 2008): 35941-35953.
PMID
18945674
Source
pubmed
Published In
The Journal of biological chemistry
Volume
283
Issue
51
Publish Date
2008
Start Page
35941
End Page
35953
DOI
10.1074/jbc.M804543200

Targeting N-cadherin enhances antitumor activity of cytotoxic therapies in melanoma treatment.

Malignant transformation in melanoma is characterized by a phenotype "switch" from E- to N-cadherin, which is associated with increased motility and invasiveness of the tumor and altered signaling, leading to decreased apoptosis. We hypothesized that the novel pentapeptide (ADH-1), which disrupts N-cadherin adhesion, could sensitize melanoma tumors to the cytotoxic effects of chemotherapy. N-cadherin-expressing human melanoma-derived cell lines were used to generate xenografts in animal models to study isolated limb infusion with melphalan and systemic chemotherapy with temozolomide. We report here that melphalan in combination with ADH-1 significantly reduced tumor growth up to 30-fold over melphalan alone. ADH-1 enhancement of response to melphalan was associated with increased formation of DNA adducts, increased apoptosis, and intracellular signaling changes associated with focal adhesions and fibroblast growth factor receptors. Targeted therapy using an N-cadherin antagonist can dramatically augment the antitumor effects of chemotherapy and is a novel approach to optimizing treatment for melanoma.

Authors
Augustine, CK; Yoshimoto, Y; Gupta, M; Zipfel, PA; Selim, MA; Febbo, P; Pendergast, AM; Peters, WP; Tyler, DS
MLA Citation
Augustine, CK, Yoshimoto, Y, Gupta, M, Zipfel, PA, Selim, MA, Febbo, P, Pendergast, AM, Peters, WP, and Tyler, DS. "Targeting N-cadherin enhances antitumor activity of cytotoxic therapies in melanoma treatment." Cancer Res 68.10 (May 15, 2008): 3777-3784.
PMID
18483261
Source
pubmed
Published In
Cancer Research
Volume
68
Issue
10
Publish Date
2008
Start Page
3777
End Page
3784
DOI
10.1158/0008-5472.CAN-07-5949

Abl tyrosine kinases modulate cadherin-dependent adhesion upstream and downstream of Rho family GTPases.

Formation and dissolution of intercellular adhesions are processes of paramount importance during tissue morphogenesis and for pathological conditions such as tumor metastasis. Cadherin-mediated intercellular adhesion requires dynamic regulation of the actin cytoskeleton. The pathways that link cadherin signaling to cytoskeletal regulation remain poorly defined. We have recently uncovered a novel role for the Abl family of tyrosine kinases linking cadherin-mediated adhesion to actin dynamics via the regulation of Rho family GTPases. Abl kinases are activated by cadherin engagement, localize to cell-cell junctions and are required for the formation of adherens junctions. Notably, we showed that Abl kinases are required for Rac activation during formation of adherens junctions, and also regulate a Rho-ROCK-myosin signaling pathway that is required for the maintenance of intercellular adhesion. Here we show that Abl kinases regulate the formation and strengthening of adherens junctions downstream of active Rac, and that Abl tyrosine kinases are components of a positive feed-back loop that employs the Crk/CrkL adaptor proteins to promote the formation and maturation of adherens junctions.

Authors
Zandy, NL; Pendergast, AM
MLA Citation
Zandy, NL, and Pendergast, AM. "Abl tyrosine kinases modulate cadherin-dependent adhesion upstream and downstream of Rho family GTPases." Cell Cycle 7.4 (February 15, 2008): 444-448. (Review)
PMID
18235247
Source
pubmed
Published In
Cell Cycle
Volume
7
Issue
4
Publish Date
2008
Start Page
444
End Page
448
DOI
10.4161/cc.7.4.5452

Phosphorylation of c-Abl by protein kinase Pak2 regulates differential binding of ABI2 and CRK

The tyrosine kinase c-Abl is implicated in a variety of cellular processes that are tightly regulated by c-Abl kinase activity and/or by interactions between c-Abl and other signaling molecules. The interaction of c-Abl with the Abl interactor protein Abi2 is shown to be negatively regulated by phosphorylation of serines 637 and 638. These serines are adjacent to the PxxP motif (PTPPKRS637S638SFR) that binds the SH3 domain of Abi. Phosphorylation of the Abl 593-730 fragment by Pak2 dramatically reduces Abi2 binding (∼90%). Mutation of serines 637-639 to alanine (3A) or aspartate (3D) results in an increased tyrosine kinase activity of c-Abl 3D, and a slight reduction of the activity of the 3A mutant, as compared to wild-type (WT) c-Abl. The interaction between Abi2 and c-Abl 3D is inhibited by 80%, as compared to WT c-Abl or c-Abl 3A. This is accompanied by a 2-fold increase in binding of Crk to c-Abl 3D. The data indicate a molecular mechanism whereby phosphorylation of c-Abl by Pak2 inhibits the interaction between the SH3 domain of Abi2 and the PxxP motif of c-Abl. This phosphorylation enhances the association of c-Abl with the substrate Crk and increases c-Abl-mediated phosphorylation of Crk, thus altering the association of Crk with other signaling molecules. © 2008 American Chemical Society.

Authors
Jung, J-H; Pendergast, AM; Zipfel, PA; Traugh, JA
MLA Citation
Jung, J-H, Pendergast, AM, Zipfel, PA, and Traugh, JA. "Phosphorylation of c-Abl by protein kinase Pak2 regulates differential binding of ABI2 and CRK." Biochemistry 47.3 (2008): 1094-1104.
PMID
18161990
Source
scival
Published In
Biochemistry
Volume
47
Issue
3
Publish Date
2008
Start Page
1094
End Page
1104
DOI
10.1021/bi701533j

Defective T cell development and function in the absence of Abelson kinases.

Thymocyte proliferation, survival, and differentiation are tightly controlled by signaling from the pre-TCR. In this study, we show for the first time that the Abelson (Abl) kinases regulate proximal signaling downstream of the pre-TCR. Conditional deletion of Abl kinases in thymocytes reveals a cell-autonomous role for these proteins in T cell development. The conditional knockout mice have reduced numbers of thymocytes, exhibit an increase in the percentage of the CD4(-)CD8(-) double-negative population, and are partially blocked in the transition to the CD4(+)CD8(+) double-positive stage. Moreover, the total number of T cells is greatly reduced in the Abl mutant mice, and the null T cells exhibit impaired TCR-induced signaling, proliferation, and cytokine production. Notably, Abl mutant mice are compromised in their ability to produce IFN-positive CD8 T cells and exhibit impaired CD8(+) T cell expansion in vivo upon Listeria monocytogenes infection. Furthermore, Ab production in response to T cell-dependent Ag is severely impaired in the Abl mutant mice. Together these findings reveal cell-autonomous roles for the Abl family kinases in both T cell development and mature T cell function, and show that loss of these kinases specifically in T cells results in compromised immunity.

Authors
Gu, JJ; Zhang, N; He, Y-W; Koleske, AJ; Pendergast, AM
MLA Citation
Gu, JJ, Zhang, N, He, Y-W, Koleske, AJ, and Pendergast, AM. "Defective T cell development and function in the absence of Abelson kinases." J Immunol 179.11 (December 1, 2007): 7334-7343.
PMID
18025176
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
179
Issue
11
Publish Date
2007
Start Page
7334
End Page
7343

Abl tyrosine kinases regulate cell-cell adhesion through Rho GTPases.

Adherens junctions are calcium-dependent cell-cell contacts that link neighboring cells through cadherin receptors. Coordinated regulation of the actin cytoskeleton by the Rho GTPases is required for the formation and dissolution of adherens junctions. However, the pathways that link cadherin signaling to cytoskeletal regulation remain poorly defined. Here we identify the Abl family kinases as critical mediators of cadherin-mediated adhesion. Endogenous Abl family kinases, Abl and Arg, are activated and required for Rac activation after cadherin engagement and regulate the formation and maintenance of adherens junctions in mammalian cells. Significantly, we show that Abl-dependent regulation of the Rho-ROCK-myosin signaling pathway is critical for the maintenance of adherens junctions. Inhibition of the Abl kinases in epithelial sheets results in the activation of Rho and its downstream target ROCK, leading to enhanced phosphorylation of the myosin regulatory light chain. These signaling events result in enhanced stress fiber formation and increased actomyosin contractility, thereby disrupting adherens junctions. Conversely, Arg gain of function promotes adherens junction formation through a Crk-dependent pathway in cells with weak junctions. These data identify the Abl kinases as a regulatory link between the cadherin-catenin adhesion complex and the actin cytoskeleton through regulation of Rac and Rho during adherens junction formation, and also reveal a functional link between Abl and Rho that is essential for adherens junction stability.

Authors
Zandy, NL; Playford, M; Pendergast, AM
MLA Citation
Zandy, NL, Playford, M, and Pendergast, AM. "Abl tyrosine kinases regulate cell-cell adhesion through Rho GTPases." Proc Natl Acad Sci U S A 104.45 (November 6, 2007): 17686-17691.
PMID
17965237
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
104
Issue
45
Publish Date
2007
Start Page
17686
End Page
17691
DOI
10.1073/pnas.0703077104

Abi-1 forms an epidermal growth factor-inducible complex with Cbl: role in receptor endocytosis.

The Abl-interactor (Abi) proteins are involved in the regulation of actin polymerization and have recently been shown to modulate epidermal growth factor receptor (EGFR) endocytosis. Here we describe the identification of a novel complex between Abi-1 and the Cbl ubiquitin ligase that is induced by stimulation with EGF. Notably, an Abi-1 mutant lacking the SH3 domain (DeltaSH3) fails to interact with Cbl and inhibits EGFR internalization. We show that expression of the Abi-1DeltaSH3 mutant inhibits Cbl accumulation at the plasma membrane after EGF treatment. We have previously shown that the oncogenic Abl tyrosine kinase inhibits EGFR internalization. Here we report that the oncogenic Abl kinase disrupts the EGF-inducible Abi-1/Cbl complex, highlighting the importance of Abl kinases and downstream effectors in the regulation of EGFR internalization. Thus, our work reveals a new role for oncogenic Abl tyrosine kinases in the regulation of the Abi-1/Cbl protein complex and uncovers a role for the Abi-1/Cbl complex in the regulation of EGFR endocytosis.

Authors
Tanos, BE; Pendergast, AM
MLA Citation
Tanos, BE, and Pendergast, AM. "Abi-1 forms an epidermal growth factor-inducible complex with Cbl: role in receptor endocytosis." Cell Signal 19.7 (July 2007): 1602-1609.
PMID
17395426
Source
pubmed
Published In
Cellular Signalling
Volume
19
Issue
7
Publish Date
2007
Start Page
1602
End Page
1609
DOI
10.1016/j.cellsig.2007.02.008

Abl tyrosine kinase regulates endocytosis of the epidermal growth factor receptor.

Signal attenuation from ligand-activated epidermal growth factor receptor (EGFR) is mediated in part by receptor endocytosis and trafficking to the lysosomal degradative compartment. Uncoupling the activated EGFR from endocytosis and degradation has emerged as a mechanism for oncogenic activation of the EGFR. The Abl nonreceptor tyrosine kinase is activated by ligand-stimulated EGFR, but the role of Abl in EGFR signaling has not been defined. Here we uncovered a novel role for the activated Abl kinase in the regulation of EGFR endocytosis. We show that activated Abl impairs EGFR internalization. Moreover, we show that activated Abl phosphorylates the EGFR primarily on tyrosine 1173, and that mutation of this site to phenylalanine restores ligand-dependent endocytosis of the EGFR in the presence of activated Abl. Furthermore, we show that activated Abl allows the ligand-activated EGFR to escape Cbl-dependent down-regulation by inhibiting the accumulation of Cbl at the plasma membrane in response to epidermal growth factor stimulation and disrupting the formation of the EGFR.Cbl complex without affecting Cbl protein stability. These findings reveal a novel role for Abl in promoting increased cell-surface expression of the EGFR and suggest that Abl/EGFR signaling may cooperate in human tumors.

Authors
Tanos, B; Pendergast, AM
MLA Citation
Tanos, B, and Pendergast, AM. "Abl tyrosine kinase regulates endocytosis of the epidermal growth factor receptor." J Biol Chem 281.43 (October 27, 2006): 32714-32723.
PMID
16943190
Source
pubmed
Published In
The Journal of biological chemistry
Volume
281
Issue
43
Publish Date
2006
Start Page
32714
End Page
32723
DOI
10.1074/jbc.M603126200

The Caenorhabditis elegans ABL-1 tyrosine kinase is required for Shigella flexneri pathogenesis.

Shigellosis is a diarrheal disease caused by the gram-negative bacterium Shigella flexneri. Following ingestion of the bacterium, S. flexneri interferes with innate immunity, establishes an infection within the human colon, and initiates an inflammatory response that results in destruction of the tissue lining the gut. Examination of host cell factors required for S. flexneri pathogenesis in vivo has proven difficult due to limited host susceptibility. Here we report the development of a pathogenesis system that involves the use of Caenorhabditis elegans as a model organism to study S. flexneri virulence determinants and host molecules required for pathogenesis. We show that S. flexneri-mediated killing of C. elegans correlates with bacterial accumulation in the intestinal tract of the animal. The S. flexneri virulence plasmid, which encodes a type III secretory system as well as various virulence determinants crucial for pathogenesis in mammalian systems, was found to be required for maximal C. elegans killing. Additionally, we demonstrate that ABL-1, the C. elegans homolog of the mammalian c-Abl nonreceptor tyrosine kinase ABL1, is required for S. flexneri pathogenesis in nematodes. These data demonstrate the feasibility of using C. elegans to study S. flexneri pathogenesis in vivo and provide insight into host factors that contribute to S. flexneri pathogenesis.

Authors
Burton, EA; Pendergast, AM; Aballay, A
MLA Citation
Burton, EA, Pendergast, AM, and Aballay, A. "The Caenorhabditis elegans ABL-1 tyrosine kinase is required for Shigella flexneri pathogenesis." Appl Environ Microbiol 72.7 (July 2006): 5043-5051.
PMID
16820504
Source
pubmed
Published In
Applied and environmental microbiology
Volume
72
Issue
7
Publish Date
2006
Start Page
5043
End Page
5051
DOI
10.1128/AEM.00558-06

Regulation of the interactions of c-Abl with Abi2 and Crk by Pak2-mediated phosphorylation

Authors
Jung, JH; Pendergast, AM; Zipfel, PA; Traugh, JA
MLA Citation
Jung, JH, Pendergast, AM, Zipfel, PA, and Traugh, JA. "Regulation of the interactions of c-Abl with Abi2 and Crk by Pak2-mediated phosphorylation." March 6, 2006.
Source
wos-lite
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
20
Issue
4
Publish Date
2006
Start Page
A497
End Page
A497

Role for the Abi/wave protein complex in T cell receptor-mediated proliferation and cytoskeletal remodeling.

BACKGROUND: The molecular reorganization of signaling molecules after T cell receptor (TCR) activation is accompanied by polymerization of actin at the site of contact between a T cell and an antigen-presenting cell (APC), as well as extension of actin-rich lamellipodia around the APC. Actin polymerization is critical for the fidelity and efficiency of the T cell response to antigen. The ability of T cells to polymerize actin is critical for several steps in T cell activation including TCR clustering, mature immunological synapse formation, calcium flux, IL-2 production, and proliferation. Activation of the Rac GTPase has been linked to regulation of actin polymerization after TCR stimulation. However, the molecules required for TCR-mediated actin polymerization downstream of activated Rac have remained elusive. Here we identify a novel role for the Abi/Wave protein complex, which signals downstream of activated Rac, in the regulation of actin polymerization and T cell activation in response to TCR stimulation. RESULTS: Here we show that Abi and Wave rapidly translocate from the T cell cytoplasm to the T cell:B cell contact site in the presence of antigen. Abi and Wave colocalize with actin at the T cell:B cell conjugation site. Moreover, Wave and Abi are necessary for actin polymerization after T cell activation, and loss of Abi proteins in mice impairs TCR-induced cell proliferation and IL-2 production in primary T cells. Significantly, the impairment in actin polymerization in cells lacking Abi proteins is due to the inability of Wave proteins to localize to the T cell:B cell contact site in the presence of antigen, rather than the destabilization of the components of the Wave protein complex. CONCLUSIONS: The Abi/Wave complex is a novel regulator of TCR-mediated actin dynamics, IL-2 production, and proliferation.

Authors
Zipfel, PA; Bunnell, SC; Witherow, DS; Gu, JJ; Chislock, EM; Ring, C; Pendergast, AM
MLA Citation
Zipfel, PA, Bunnell, SC, Witherow, DS, Gu, JJ, Chislock, EM, Ring, C, and Pendergast, AM. "Role for the Abi/wave protein complex in T cell receptor-mediated proliferation and cytoskeletal remodeling." Curr Biol 16.1 (January 10, 2006): 35-46.
PMID
16401422
Source
pubmed
Published In
Current Biology
Volume
16
Issue
1
Publish Date
2006
Start Page
35
End Page
46
DOI
10.1016/j.cub.2005.12.024

Lower capillarization, VEGF protein, and VEGF mRNA response to acute exercise in the vastus lateralis muscle of aged vs. young women.

In humans, the majority of studies demonstrate an age-associated reduction in the number of capillaries surrounding skeletal muscle fibers; however, recent reports in rats suggest that muscle capillarization is well maintained with advanced age. In sedentary and trained men, aging lowers the number of capillaries surrounding type II, but not type I, skeletal muscle fibers. The fiber type-specific effect of aging on muscle capillarization is unknown in women. Vascular endothelial growth factor (VEGF) is important in the basal maintenance of skeletal muscle capillarization, and lower VEGF expression is associated with increased age in nonskeletal muscle tissue of women. Compared with young women (YW), we hypothesized that aged women (AW) would demonstrate 1) lower muscle capillarization in a fiber type-specific manner and 2) lower VEGF and VEGF receptor expression at rest and in response to acute exercise. Nine sedentary AW (70 + 8 yr) and 11 YW (22 + 3 yr) had vastus lateralis muscle biopsies obtained before and at 4 h after a submaximal exercise bout for the measurement of morphometry and VEGF and VEGF receptor expression. In AW compared with YW, muscle capillary contacts were lower overall (YW: 2.36 + 0.32 capillaries; AW: 2.08 + 0.17 capillaries), specifically in type II (YW: 2.37 + 0.39 capillaries; AW: 1.91 + 0.36 capillaries) but not type I fibers (YW: 2.36 + 0.34 capillaries; AW: 2.26 + 0.24 capillaries). Muscle VEGF protein was 35% lower at rest, and the exercise-induced increase in VEGF mRNA was 50% lower in AW compared with YW. There was no effect of age on VEGF receptor expression. These results provide evidence that, in the vastus lateralis of women, 1) capillarization surrounding type II muscle fibers is lower in AW compared with YW and 2) resting VEGF protein and the VEGF mRNA response to exercise are lower in AW compared with YW.

Authors
Croley, AN; Zwetsloot, KA; Westerkamp, LM; Ryan, NA; Pendergast, AM; Hickner, RC; Pofahl, WE; Gavin, TP
MLA Citation
Croley, AN, Zwetsloot, KA, Westerkamp, LM, Ryan, NA, Pendergast, AM, Hickner, RC, Pofahl, WE, and Gavin, TP. "Lower capillarization, VEGF protein, and VEGF mRNA response to acute exercise in the vastus lateralis muscle of aged vs. young women." J Appl Physiol (1985) 99.5 (November 2005): 1872-1879.
PMID
16024519
Source
pubmed
Published In
Journal of applied physiology (Bethesda, Md. : 1985)
Volume
99
Issue
5
Publish Date
2005
Start Page
1872
End Page
1879
DOI
10.1152/japplphysiol.00498.2005

Abl kinases regulate actin comet tail elongation via an N-WASP-dependent pathway.

Microbial pathogens have evolved diverse strategies to modulate the host cell cytoskeleton to achieve a productive infection and have proven instrumental for unraveling the molecular machinery that regulates actin polymerization. Here we uncover a mechanism for Shigella flexneri-induced actin comet tail elongation that links Abl family kinases to N-WASP-dependent actin polymerization. We show that the Abl kinases are required for Shigella actin comet tail formation, maximal intracellular motility, and cell-to-cell spread. Abl phosphorylates N-WASP, a host cell protein required for actin comet tail formation, and mutation of the Abl phosphorylation sites on N-WASP impairs comet tail elongation. Furthermore, we show that defective comet tail formation in cells lacking Abl kinases is rescued by activated forms of N-WASP. These data demonstrate for the first time that the Abl kinases play a role in the intracellular motility and intercellular dissemination of Shigella and uncover a new role for Abl kinases in the regulation of pathogen motility.

Authors
Burton, EA; Oliver, TN; Pendergast, AM
MLA Citation
Burton, EA, Oliver, TN, and Pendergast, AM. "Abl kinases regulate actin comet tail elongation via an N-WASP-dependent pathway." Mol Cell Biol 25.20 (October 2005): 8834-8843.
PMID
16199863
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
25
Issue
20
Publish Date
2005
Start Page
8834
End Page
8843
DOI
10.1128/MCB.25.20.8834-8843.2005

Stress and death: breaking up the c-Abl/14-3-3 complex in apoptosis.

Authors
Pendergast, AM
MLA Citation
Pendergast, AM. "Stress and death: breaking up the c-Abl/14-3-3 complex in apoptosis." Nat Cell Biol 7.3 (March 2005): 213-214.
PMID
15738971
Source
pubmed
Published In
Nature Cell Biology
Volume
7
Issue
3
Publish Date
2005
Start Page
213
End Page
214
DOI
10.1038/ncb0305-213

ABI2-deficient mice exhibit defective cell migration, aberrant dendritic spine morphogenesis, and deficits in learning and memory.

The Abl-interactor (Abi) family of adaptor proteins has been linked to signaling pathways involving the Abl tyrosine kinases and the Rac GTPase. Abi proteins localize to sites of actin polymerization in protrusive membrane structures and regulate actin dynamics in vitro. Here we demonstrate that Abi2 modulates cell morphogenesis and migration in vivo. Homozygous deletion of murine abi2 produced abnormal phenotypes in the eye and brain, the tissues with the highest Abi2 expression. In the absence of Abi2, secondary lens fiber orientation and migration were defective in the eye, without detectable defects in proliferation, differentiation, or apoptosis. These phenotypes were consistent with the localization of Abi2 at adherens junctions in the developing lens and at nascent epithelial cell adherens junctions in vitro. Downregulation of Abi expression by RNA interference impaired adherens junction formation and correlated with downregulation of the Wave actin-nucleation promoting factor. Loss of Abi2 also resulted in cell migration defects in the neocortex and hippocampus, abnormal dendritic spine morphology and density, and severe deficits in short- and long-term memory. These findings support a role for Abi2 in the regulation of cytoskeletal dynamics at adherens junctions and dendritic spines, which is critical for intercellular connectivity, cell morphogenesis, and cognitive functions.

Authors
Grove, M; Demyanenko, G; Echarri, A; Zipfel, PA; Quiroz, ME; Rodriguiz, RM; Playford, M; Martensen, SA; Robinson, MR; Wetsel, WC; Maness, PF; Pendergast, AM
MLA Citation
Grove, M, Demyanenko, G, Echarri, A, Zipfel, PA, Quiroz, ME, Rodriguiz, RM, Playford, M, Martensen, SA, Robinson, MR, Wetsel, WC, Maness, PF, and Pendergast, AM. "ABI2-deficient mice exhibit defective cell migration, aberrant dendritic spine morphogenesis, and deficits in learning and memory." Mol Cell Biol 24.24 (December 2004): 10905-10922.
PMID
15572692
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
24
Issue
24
Publish Date
2004
Start Page
10905
End Page
10922
DOI
10.1128/MCB.24.24.10905-10922.2004

Requirement for Abl kinases in T cell receptor signaling.

BACKGROUND: The c-Abl and Arg proteins comprise a unique family of nonreceptor tyrosine kinases that have been implicated in the regulation of cell proliferation and survival, cytoskeletal reorganization, cell migration, and the response to oxidative stress and DNA damage. Targeted deletion or mutation of c-Abl in mice results in a variety of immune system phenotypes, including splenic and thymic atrophy, lymphopenia, and an increased susceptibility to infection. However, despite the generation of these mice over a decade ago, little is known regarding the mechanisms responsible for these phenotypes or the immune-related consequences of ablation of both the c-Abl and Arg kinases, which are coexpressed in lymphoid tissues. RESULTS: Here, we report that T cell receptor (TCR) stimulation results in activation of the endogenous Abl kinases. We demonstrate that Zap70 and the transmembrane adaptor linker for activation of T cells (LAT) are targets of the Abl kinases, and that loss of Abl kinase activity reduces TCR-induced Zap70 phosphorylation at tyrosine 319. This correlates with diminished LAT tyrosine phosphorylation, as well as reduced tyrosine phosphorylation and recruitment of phospholipase Cgamma1 to LAT. Significantly, we show that Abl kinase activity is required for maximal signaling leading to transcription of the IL-2 promoter, as well as TCR-induced IL-2 production and proliferation of primary T cells. CONCLUSIONS: We conclude that the Abl kinases have a role in the regulation of TCR-mediated signal transduction leading to IL-2 production and cell proliferation.

Authors
Zipfel, PA; Zhang, W; Quiroz, M; Pendergast, AM
MLA Citation
Zipfel, PA, Zhang, W, Quiroz, M, and Pendergast, AM. "Requirement for Abl kinases in T cell receptor signaling." Curr Biol 14.14 (July 27, 2004): 1222-1231.
PMID
15268851
Source
pubmed
Published In
Current Biology
Volume
14
Issue
14
Publish Date
2004
Start Page
1222
End Page
1231
DOI
10.1016/j.cub.2004.07.021

Abl interactor 1 (Abi-1) wave-binding and SNARE domains regulate its nucleocytoplasmic shuttling, lamellipodium localization, and wave-1 levels.

The Abl interactor 1 (Abi-1) protein has been implicated in the regulation of actin dynamics and localizes to the tips of lamellipodia and filopodia. Here, we show that Abi-1 binds the actin nucleator protein Wave-1 through an amino-terminal Wave-binding (WAB) domain and that disruption of the Abi-1-Wave-1 interaction prevents Abi-1 from reaching the tip of the lamellipodium. Abi-1 binds to the Wave homology domain of Wave-1, a region that is required for translocation of Wave-1 to the lamellipodium. Mouse embryo fibroblasts that lack one allele of Abi-1 and are homozygous null for the related Abi-2 protein exhibit decreased Wave-1 protein levels. This phenotype is rescued by Abi-1 proteins that retain Wave-1 binding but not by Abi-1 mutants that cannot bind to Wave-1. Moreover, we uncovered an overlapping SNARE domain in the amino terminus of Abi-1 that interacts with Syntaxin-1, a SNARE family member. Further, we demonstrated that Abi-1 shuttles in and out of the nucleus in a leptomycin B (LMB)-dependent manner and that complete nuclear translocation of Abi-1 in the absence of LMB requires the combined inactivation of the SNARE, WAB, and SH3 domains of Abi-1. Thus, Abi-1 undergoes nucleocytoplasmic shuttling and functions at the leading edge to regulate Wave-1 localization and protein levels.

Authors
Echarri, A; Lai, MJ; Robinson, MR; Pendergast, AM
MLA Citation
Echarri, A, Lai, MJ, Robinson, MR, and Pendergast, AM. "Abl interactor 1 (Abi-1) wave-binding and SNARE domains regulate its nucleocytoplasmic shuttling, lamellipodium localization, and wave-1 levels." Mol Cell Biol 24.11 (June 2004): 4979-4993.
PMID
15143189
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
24
Issue
11
Publish Date
2004
Start Page
4979
End Page
4993
DOI
10.1128/MCB.24.11.4979-4993.2004

Bidirectional signaling links the Abelson kinases to the platelet-derived growth factor receptor.

The c-Abl nonreceptor tyrosine kinase is activated by growth factor signals such as the platelet-derived growth factor (PDGF) and functions downstream of the PDGF-beta receptor (PDGFR) to mediate biological processes such as membrane ruffling, mitogenesis, and chemotaxis. Here, we show that the related kinase Arg is activated downstream of PDGFRs in a manner dependent on Src family kinases and phospholipase C gamma1 (PLC-gamma1)-mediated phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis, as we showed previously for c-Abl. PIP2, a highly abundant phosphoinositide known to regulate cytoskeletal and membrane proteins, inhibits the tyrosine kinase activities of both Arg and c-Abl in vitro and in cells. We now demonstrate that c-Abl and Arg form inducible complexes with and are phosphorylated by the PDGFR tyrosine kinase in vitro and in vivo. Moreover, c-Abl and Arg, in turn, phosphorylate the PDGFR. We show that c-Abl and Arg exhibit nonredundant functions downstream of the activated PDGFR. Reintroduction of c-Abl into Arg-Abl double-null fibroblasts rescues the ability of PLC-gamma1 to increase PDGF-mediated chemotaxis, while reexpression of Arg fails to rescue the chemotaxis defect. These data show that, although both kinases are activated and form complexes with proteins in the PDGFR signaling pathway, only c-Abl functions downstream of PLC-gamma1 to mediate chemotaxis.

Authors
Plattner, R; Koleske, AJ; Kazlauskas, A; Pendergast, AM
MLA Citation
Plattner, R, Koleske, AJ, Kazlauskas, A, and Pendergast, AM. "Bidirectional signaling links the Abelson kinases to the platelet-derived growth factor receptor." Mol Cell Biol 24.6 (March 2004): 2573-2583.
PMID
14993293
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
24
Issue
6
Publish Date
2004
Start Page
2573
End Page
2583

Elevated expression of a subset of interferon inducible genes in primary bone marrow cells expressing p185 Bcr-Abl versus p210 Bcr-Abl by DNA microarray analysis.

p185 Bcr-Abl has a more aggressive biological/clinical leukemia phenotype than p210 Bcr-Abl. In this study, we examined differential gene expression using microarrays to determine if upregulation or downregulation of specific genes may explain the distinct phenotypes produced by the two Bcr-Abl forms. RNA was collected from mouse bone marrow mononuclear cells expressing equivalent levels of p185 or p210, and the RNAs were subjected to microarray analysis. Significant differences in gene expression were observed on hierarchical clustering. A group of interferon-gamma-inducible genes, including those encoding a family of 47 kDa GTPases, were significantly increased in p185 versus p210. This family of GTPases has previously been implicated in interferon-gamma-induced resistance against intracellular pathogens, however their exact cellular functions are unknown. Our data suggest that their increased expression may contribute to the biological/clinical phenotype associated with p185.

Authors
Advani, AS; Dressman, HK; Quiroz, M; Taylor, GA; Pendergast, AM
MLA Citation
Advani, AS, Dressman, HK, Quiroz, M, Taylor, GA, and Pendergast, AM. "Elevated expression of a subset of interferon inducible genes in primary bone marrow cells expressing p185 Bcr-Abl versus p210 Bcr-Abl by DNA microarray analysis." Leuk Res 28.3 (March 2004): 285-294.
PMID
14687624
Source
pubmed
Published In
Leukemia Research
Volume
28
Issue
3
Publish Date
2004
Start Page
285
End Page
294

Abl tyrosine kinases are required for infection by Shigella flexneri.

Infection by the opportunistic bacterial pathogen Shigella flexneri stimulates tyrosine phosphorylation of host cell proteins, but the kinases involved and their effects on the regulation of cell signaling pathways during bacterial entry remain largely undefined. Here, we demonstrate a requirement for the Abl family of tyrosine kinases during Shigella internalization. Family members Abl and Arg are catalytically activated upon Shigella infection, accumulate at the site of bacterial entry, and are required for efficient bacterial uptake, as internalization is blocked upon targeted deletion of these kinases or treatment with a specific pharmacological inhibitor. We identify the adapter protein Crk as a target for Abl kinases during Shigella uptake, and show that a phosphorylation-deficient Crk mutant significantly inhibits bacterial uptake. Moreover, we define a novel signaling pathway activated during Shigella entry that links Abl kinase phosphorylation of Crk to activation of the Rho family GTPases Rac and Cdc42. Together, these findings reveal a new role for the Abl kinases, and suggest a novel approach to treatment of Shigella infections through inhibition of host cell signaling pathways.

Authors
Burton, EA; Plattner, R; Pendergast, AM
MLA Citation
Burton, EA, Plattner, R, and Pendergast, AM. "Abl tyrosine kinases are required for infection by Shigella flexneri." EMBO J 22.20 (October 15, 2003): 5471-5479.
PMID
14532119
Source
pubmed
Published In
EMBO Journal
Volume
22
Issue
20
Publish Date
2003
Start Page
5471
End Page
5479
DOI
10.1093/emboj/cdg512

Postsynaptic requirement for Abl kinases in assembly of the neuromuscular junction.

Agrin signals through the muscle-specific receptor tyrosine kinase (MuSK) to cluster acetylcholine receptors (AChRs) on the postsynaptic membrane of the neuromuscular junction (NMJ). This stands as the prevailing model of synapse induction by a presynaptic factor, yet the agrin-dependent MuSK signaling cascade is largely undefined. Abl1 (previously known as Abl) and the Abl1-related gene product Abl2 (previously known as Arg) define a family of tyrosine kinases that regulate actin structure and presynaptic axon guidance. Here we show that the Abl kinases are critical mediators of postsynaptic assembly downstream of agrin and MuSK. In mouse muscle, Abl kinases were localized to the postsynaptic membrane of the developing NMJ. In cultured myotubes, Abl kinase activity was required for agrin-induced AChR clustering and enhancement of MuSK tyrosine phosphorylation. Moreover, MuSK and Abl kinases effected reciprocal tyrosine phosphorylation and formed a complex after agrin engagement. Our findings suggest that Abl kinases provide the developing synapse with the kinase activity required for signal amplification and the intrinsic cytoskeletal regulatory capacity required for assembly and remodeling.

Authors
Finn, AJ; Feng, G; Pendergast, AM
MLA Citation
Finn, AJ, Feng, G, and Pendergast, AM. "Postsynaptic requirement for Abl kinases in assembly of the neuromuscular junction." Nat Neurosci 6.7 (July 2003): 717-723.
PMID
12796783
Source
pubmed
Published In
Nature Neuroscience
Volume
6
Issue
7
Publish Date
2003
Start Page
717
End Page
723
DOI
10.1038/nn1071

Activation and signaling of the Abl tyrosine kinase: bidirectional link with phosphoinositide signaling.

Authors
Plattner, R; Pendergast, AM
MLA Citation
Plattner, R, and Pendergast, AM. "Activation and signaling of the Abl tyrosine kinase: bidirectional link with phosphoinositide signaling." Cell Cycle 2.4 (July 2003): 273-274. (Review)
PMID
12851468
Source
pubmed
Published In
Cell Cycle
Volume
2
Issue
4
Publish Date
2003
Start Page
273
End Page
274

A new link between the c-Abl tyrosine kinase and phosphoinositide signalling through PLC-gamma1.

The c-Abl tyrosine (Tyr) kinase is activated after platelet-derived-growth factor receptor (PDGFR) stimulation in a manner that is partially dependent on Src kinase activity. However, the activity of Src kinases alone is not sufficient for activation of c-Abl by PDGFR. Here we show that functional phospholipase C-gamma1 (PLC-gamma1) is required for c-Abl activation by PDGFR. Decreasing cellular levels of phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) by PLC-gamma1-mediated hydrolysis or dephosphorylation by an inositol polyphosphate 5-phosphatase (Inp54) results in increased Abl kinase activity. c-Abl functions downstream of PLC-gamma1, as expression of kinase-inactive c-Abl blocks PLC-gamma1-induced chemotaxis towards PDGF-BB. PLC-gamma1 and c-Abl form a complex in cells that is enhanced by PDGF stimulation. After activation, c-Abl phosphorylates PLC-gamma1 and negatively modulates its function in vivo. These findings uncover a newly discovered functional interdependence between non-receptor Tyr kinase and lipid signalling pathways.

Authors
Plattner, R; Irvin, BJ; Guo, S; Blackburn, K; Kazlauskas, A; Abraham, RT; York, JD; Pendergast, AM
MLA Citation
Plattner, R, Irvin, BJ, Guo, S, Blackburn, K, Kazlauskas, A, Abraham, RT, York, JD, and Pendergast, AM. "A new link between the c-Abl tyrosine kinase and phosphoinositide signalling through PLC-gamma1." Nat Cell Biol 5.4 (April 2003): 309-319.
PMID
12652307
Source
pubmed
Published In
Nature Cell Biology
Volume
5
Issue
4
Publish Date
2003
Start Page
309
End Page
319
DOI
10.1038/ncb949

Erratum: Abl tyrosine kinases are required for infection by Shigella flexneri (EMBO Journal (2003) 22 (5471-5479))

Authors
Burton, EA; Plattner, R; Pendergast, AM
MLA Citation
Burton, EA, Plattner, R, and Pendergast, AM. "Erratum: Abl tyrosine kinases are required for infection by Shigella flexneri (EMBO Journal (2003) 22 (5471-5479))." EMBO Journal 22.21 (2003): 5962--.
Source
scival
Published In
EMBO Journal
Volume
22
Issue
21
Publish Date
2003
Start Page
5962-
DOI
10.1093/emboj/cdg587

Elevated expression of numerous interferon inducible genes in primary bone marrow cells expressing p185 Bcr-Abl versus p210 Bcr-Abl by DNA microarray analysis.

Authors
Advani, AS; Dressman, HK; Quiroz, M; Taylor, GA; Pendergast, AM
MLA Citation
Advani, AS, Dressman, HK, Quiroz, M, Taylor, GA, and Pendergast, AM. "Elevated expression of numerous interferon inducible genes in primary bone marrow cells expressing p185 Bcr-Abl versus p210 Bcr-Abl by DNA microarray analysis." November 16, 2002.
Source
wos-lite
Published In
Blood
Volume
100
Issue
11
Publish Date
2002
Start Page
584A
End Page
584A

Bcr-Abl variants: biological and clinical aspects.

Bcr-Abl is an oncogene that arises from fusion of the Bcr gene with the c-Abl proto-oncogene. Three different Bcr-Abl variants can be formed, depending on the amount of Bcr gene included: p185, p210, and p230. The three variants are associated with distinct types of human leukemias. Examination of the signaling pathways differentially regulated by the Bcr-Abl proteins will help us gain better insight into Bcr-Abl mediated leukemogenesis.

Authors
Advani, AS; Pendergast, AM
MLA Citation
Advani, AS, and Pendergast, AM. "Bcr-Abl variants: biological and clinical aspects." Leuk Res 26.8 (August 2002): 713-720. (Review)
PMID
12191565
Source
pubmed
Published In
Leukemia Research
Volume
26
Issue
8
Publish Date
2002
Start Page
713
End Page
720

The Abl family kinases: mechanisms of regulation and signaling.

Authors
Pendergast, AM
MLA Citation
Pendergast, AM. "The Abl family kinases: mechanisms of regulation and signaling." Adv Cancer Res 85 (2002): 51-100. (Review)
PMID
12374288
Source
pubmed
Published In
Advances in cancer research
Volume
85
Publish Date
2002
Start Page
51
End Page
100

Activated c-Abl is degraded by the ubiquitin-dependent proteasome pathway.

C-Abl is a nonreceptor tyrosine kinase that is tightly regulated in the cell. Genetic data derived from studies in flies and mice strongly support a role for Abl kinases in the regulation of the cytoskeleton (reviewed in [1,2]). C-Abl can be activated by several stimuli, including oxidative stress [3], DNA damage [4], integrin engagement [5], growth factors, and Src family kinases [6]. Structural alterations elicit constitutive activation of the c-Abl tyrosine kinase, leading to oncogenic transformation. While the mechanisms that activate c-Abl are beginning to be elucidated, little is known regarding the mechanisms that downregulate activated c-Abl. Here, we show for the first time that activated c-Abl is downregulated by the ubiquitin-dependent degradation pathway. Activated forms of c-Abl are more unstable than wild-type and kinase-inactive forms. Moreover, inhibition of the 26S proteasome leads to increased c-Abl levels in vitro and in cells, and activated c-Abl proteins are ubiquitinated in vivo. Significantly, inhibition of the 26S proteasome in fibroblasts increases the levels of tyrosine-phosphorylated, endogenous c-Abl. Our data suggest a novel mechanism for irreversible downregulation of activated c-Abl, which is critical to prevent the deleterious consequences of c-Abl hyperactivation in mitogenic and cytoskeletal pathways.

Authors
Echarri, A; Pendergast, AM
MLA Citation
Echarri, A, and Pendergast, AM. "Activated c-Abl is degraded by the ubiquitin-dependent proteasome pathway." Curr Biol 11.22 (November 13, 2001): 1759-1765.
PMID
11719217
Source
pubmed
Published In
Current Biology
Volume
11
Issue
22
Publish Date
2001
Start Page
1759
End Page
1765

The Abl interactor proteins localize to sites of actin polymerization at the tips of lamellipodia and filopodia.

Cell movement is mediated by the protrusion of cytoplasm in the form of sheet- and rod-like extensions, termed lamellipodia and filopodia. Protrusion is driven by actin polymerization, a process that is regulated by signaling complexes that are, as yet, poorly defined. Since actin assembly is controlled at the tips of lamellipodia and filopodia [1], these juxtamembrane sites are likely to harbor the protein complexes that control actin polymerization dynamics underlying cell motility. An understanding of the regulation of protrusion therefore requires the characterization of the molecular components recruited to these sites. The Abl interactor (Abi) proteins, targets of Abl tyrosine kinases [2-4], have been implicated in Rac-dependent cytoskeletal reorganization in response to growth factor stimulation [5]. Here, we describe the unique localization of Abi proteins in living, motile cells. We show that Abi-1 and Abi-2b fused to enhanced yellow fluorescent protein (EYFP) are recruited to the tips of lamellipodia and filopodia. We identify the targeting domain as the homologous N terminus of these two proteins. Our findings are the first to suggest a direct involvement of members of the Abi protein family in the control of actin polymerization in protrusion events, and establish the Abi proteins as potential regulators of motility.

Authors
Stradal, T; Courtney, KD; Rottner, K; Hahne, P; Small, JV; Pendergast, AM
MLA Citation
Stradal, T, Courtney, KD, Rottner, K, Hahne, P, Small, JV, and Pendergast, AM. "The Abl interactor proteins localize to sites of actin polymerization at the tips of lamellipodia and filopodia." Curr Biol 11.11 (June 5, 2001): 891-895.
PMID
11516653
Source
pubmed
Published In
Current Biology
Volume
11
Issue
11
Publish Date
2001
Start Page
891
End Page
895

Functional interaction between c-Abl and the p21-activated protein kinase gamma-PAK.

A member of the p21-activated protein kinase (PAK) family, gamma-PAK has cytostatic properties and is activated by cellular stresses such as hyperosmolarity or DNA damage. We report herein that gamma-PAK is associated in vivo with the nonreceptor protein tyrosine kinase c-Abl. gamma-PAK phosphorylates c-Abl on sites located in the kinase domain, in a region that is implicated in protein-protein interactions and in subcellular localization. Activation of gamma-PAK in human embryonic kidney 293T cells by cotransfection with constitutively active Cdc42 induces activation of c-Abl, resulting in increased phosphotyrosine levels. Cotransfection of c-Abl and gamma-PAK elicits phosphorylation of gamma-PAK on tyrosine and down-regulation of gamma-PAK activity, promoting accumulation of inactive gamma-PAK. gamma-PAK is also phosphorylated in vitro by c-Abl. gamma-PAK activity is regulated by ubiquitination and proteolysis in vivo, as shown by immunoblotting with an anti-ubiquitin antibody in the presence of proteasome inhibitors. In summary, we describe a functional interaction between gamma-PAK and c-Abl in which gamma-PAK stimulates c-Abl tyrosine kinase activity and c-Abl phosphorylates and down-regulates gamma-PAK, suggesting the existence of a negative feedback loop between c-Abl and gamma-PAK.

Authors
Roig, J; Tuazon, PT; Zipfel, PA; Pendergast, AM; Traugh, JA
MLA Citation
Roig, J, Tuazon, PT, Zipfel, PA, Pendergast, AM, and Traugh, JA. "Functional interaction between c-Abl and the p21-activated protein kinase gamma-PAK." Proc Natl Acad Sci U S A 97.26 (December 19, 2000): 14346-14351.
PMID
11121037
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
97
Issue
26
Publish Date
2000
Start Page
14346
End Page
14351
DOI
10.1073/pnas.97.26.14346

The c-Abl tyrosine kinase is regulated downstream of the B cell antigen receptor and interacts with CD19.

c-Abl is a nonreceptor tyrosine kinase that we have recently linked to growth factor receptor signaling. The c-Abl kinase is ubiquitously expressed and localizes to the cytoplasm, plasma membrane, cytoskeleton, and nucleus. Thus, c-Abl may regulate signaling processes in multiple subcellular compartments. Targeted deletion or mutation of c-Abl in mice results in a variety of phenotypes, including splenic and thymic atrophy and lymphopenia. Additionally, lymphocytes isolated from specific compartments of c-Abl mutant mice have reduced responses to a variety of stimuli and an increased susceptibility to apoptosis following growth factor deprivation. Despite these observations, little is known regarding the signaling mechanisms responsible for these phenotypes. We report here that splenic B cells from c-Abl-deficient mice are hyporesponsive to the proliferative effects of B cell Ag receptor (BCR) stimulation. The c-Abl kinase activity and protein levels are elevated in the cytosol following activation of the BCR in B cell lines. We show that c-Abl associates with and phosphorylates the BCR coreceptor CD19, and that c-Abl and CD19 colocalize in lipid membrane rafts. These data suggest a role for c-Abl in the regulation of B cell proliferation downstream of the BCR, possibly through interactions with CD19.

Authors
Zipfel, PA; Grove, M; Blackburn, K; Fujimoto, M; Tedder, TF; Pendergast, AM
MLA Citation
Zipfel, PA, Grove, M, Blackburn, K, Fujimoto, M, Tedder, TF, and Pendergast, AM. "The c-Abl tyrosine kinase is regulated downstream of the B cell antigen receptor and interacts with CD19." J Immunol 165.12 (December 15, 2000): 6872-6879.
PMID
11120811
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
165
Issue
12
Publish Date
2000
Start Page
6872
End Page
6879

Localization and phosphorylation of Abl-interactor proteins, Abi-1 and Abi-2, in the developing nervous system.

Abl-interactor (Abi) proteins are targets of Abl-family nonreceptor tyrosine kinases and are required for Rac-dependent cytoskeletal reorganization in response to growth factor stimulation. We asked if the expression, phosphorylation, and cellular localization of Abi-1 and Abi-2 supports a role for these proteins in Abl signaling in the developing and adult mouse nervous system. In mid- to late-gestation embryos, abi-2 message is elevated in the central and peripheral nervous systems (CNS and PNS). Abi-1 mRNA is present, but not enhanced, in the CNS, and is not observed in PNS structures. Abi proteins from brain lysates undergo changes in apparent molecular weight and phosphorylation with increasing age. In the postnatal brain, abi-1 and abi-2 are expressed most prominently in cortical layers populated by projection neurons. In cultured neurons, Abi-1 and Abi-2 are concentrated in puncta throughout the cell body and processes. Both Abi and Abl proteins are present in synaptosomes and growth cone particles. Therefore, the Abi adaptors exhibit proper expression patterns and subcellular localization to participate in Abl kinase signaling in the nervous system.

Authors
Courtney, KD; Grove, M; Vandongen, H; Vandongen, A; LaMantia, AS; Pendergast, AM
MLA Citation
Courtney, KD, Grove, M, Vandongen, H, Vandongen, A, LaMantia, AS, and Pendergast, AM. "Localization and phosphorylation of Abl-interactor proteins, Abi-1 and Abi-2, in the developing nervous system." Mol Cell Neurosci 16.3 (September 2000): 244-257.
PMID
10995551
Source
pubmed
Published In
Molecular and Cellular Neuroscience
Volume
16
Issue
3
Publish Date
2000
Start Page
244
End Page
257
DOI
10.1006/mcne.2000.0865

Analysis of the biologic properties of p230 Bcr-Abl reveals unique and overlapping properties with the oncogenic p185 and p210 Bcr-Abl tyrosine kinases.

The reciprocal translocation between chromosomes 9 and 22 that fuses coding sequences of the Bcr and Abl genes is responsible for a remarkably diverse group of hematologic malignancies. A newly described 230-kd form of Bcr-Abl has been associated with an indolent myeloproliferative syndrome referred to as chronic neutrophilic leukemia. We have cloned the corresponding gene and examined the biologic and biochemical properties of p230 Bcr-Abl after retroviral-mediated gene transfer into hematopoietic cell lines and primary bone marrow cells. p230 Bcr-Abl-expressing 32D myeloid cells were fully growth factor-independent and activated similar signal transduction pathways as the well-characterized p210 and p185 forms of Bcr-Abl. In contrast, primary mouse bone marrow cells expressing p230 required exogenous hematopoietic growth factors for optimal growth, whereas p185- and p210-expressing cells were independent of growth factors. The 3 Bcr-Abl proteins exerted different effects on differentiation of bone marrow cells. p185 induced outgrowth of lymphoid precursors capable of tumor formation in immunodeficient mice. In contrast, p210- and p230-expressing bone marrow cells caused limited outgrowth of lymphoid precursors that failed to form tumors in immunodeficient mice. Removal of cytokines and autologous stroma from Bcr-Abl-expressing bone marrow cultures produced the expansion of distinct lineages by the various Bcr-Abl proteins. p185 drove expansion of cytokine-independent lymphoid progenitors, while p210 and p230 generated cytokine-independent monocyte/myeloid cells. These findings suggest that the different Bcr-Abl fusion proteins drive the expansion of different hematopoietic populations, which may explain the association of the various Bcr-Abl oncoproteins with different spectra of human leukemias. (Blood. 2000;95:2913-2921)

Authors
Quackenbush, RC; Reuther, GW; Miller, JP; Courtney, KD; Pear, WS; Pendergast, AM
MLA Citation
Quackenbush, RC, Reuther, GW, Miller, JP, Courtney, KD, Pear, WS, and Pendergast, AM. "Analysis of the biologic properties of p230 Bcr-Abl reveals unique and overlapping properties with the oncogenic p185 and p210 Bcr-Abl tyrosine kinases." Blood 95.9 (May 1, 2000): 2913-2921.
PMID
10779439
Source
pubmed
Published In
Blood
Volume
95
Issue
9
Publish Date
2000
Start Page
2913
End Page
2921

The BCR/ABL oncogene alters the chemotactic response to stromal-derived factor-1alpha.

The chemokine stromal-derived factor-1alpha (SDF-1alpha) is a chemoattractant for CD34(+) progenitor cells, in vitro and in vivo. The receptor for SDF-1alpha, CXCR-4, is a 7 transmembrane domain receptor, which is also a coreceptor for human immunodeficiency virus (HIV). Here we show that transformation of hematopoietic cell lines by BCR/ABL significantly impairs their response to SDF-1alpha. Three different hematopoietic cell lines, Ba/F3, 32Dcl3, and Mo7e, were found to express CXCR-4 and to respond to SDF-1alpha with increased migration in a transwell assay. In contrast, after transformation by the BCR/ABL oncogene, the chemotactic response to SDF-1alpha was reduced in all 3 lines. This effect was directly due to BCR/ABL, because Ba/F3 cells, in which the expression of BCR/ABL could be regulated by a tetracycline-inducible promoter, also had reduced chemotaxis to SDF-1alpha when BCR/ABL was induced. The reduced response to SDF-1alpha was not due to an inability of BCR/ABL-transformed cell lines to migrate in general, as spontaneous motility of BCR/ABL-transformed cells was increased. In mice, injection of SDF-1alpha into the spleen resulted in a transient accumulation of untransformed Ba/F3 cells, but not Ba/F3. p210(BCR/ABL) cells administered simultaneously. The mechanism may involve inhibition of CXCR-4 receptor function, because in BCR/ABL-transformed cells, CXCR-4 receptors were expressed on the cell surface, but SDF-1alpha calcium flux was inhibited. Because SDF-1alpha and CXCR-4 are felt to be involved in progenitor cell homing to marrow, the abnormality decribed here could contribute to the homing and retention defects typical of immature myeloid cells in chronic myelogenous leukemia.

Authors
Salgia, R; Quackenbush, E; Lin, J; Souchkova, N; Sattler, M; Ewaniuk, DS; Klucher, KM; Daley, GQ; Kraeft, SK; Sackstein, R; Alyea, EP; von Andrian, UH; Chen, LB; Gutierrez-Ramos, JC; Pendergast, AM; Griffin, JD
MLA Citation
Salgia, R, Quackenbush, E, Lin, J, Souchkova, N, Sattler, M, Ewaniuk, DS, Klucher, KM, Daley, GQ, Kraeft, SK, Sackstein, R, Alyea, EP, von Andrian, UH, Chen, LB, Gutierrez-Ramos, JC, Pendergast, AM, and Griffin, JD. "The BCR/ABL oncogene alters the chemotactic response to stromal-derived factor-1alpha." Blood 94.12 (December 15, 1999): 4233-4246.
PMID
10590068
Source
pubmed
Published In
Blood
Volume
94
Issue
12
Publish Date
1999
Start Page
4233
End Page
4246

c-Abl is activated by growth factors and Src family kinases and has a role in the cellular response to PDGF.

The c-Abl tyrosine kinase localizes to the cytoplasm and plasma membrane in addition to the nucleus. However, there is little information regarding a role for c-Abl in the cytoplasm/plasma membrane compartments. Here we report that a membrane pool of c-Abl is activated by the growth factors PDGF and EGF in fibroblasts. The pattern and kinetics of activation are similar to growth factor activation of Src family kinases. To determine whether a link existed between activation of c-Abl and members of the Src family, we examined c-Abl kinase activity in cells that expressed oncogenic Src proteins. We found that c-Abl kinase activity was increased by 10- to 20-fold in these cells, and that Src and Fyn kinases directly phosphorylated c-Abl in vitro. Furthermore, overexpression of wild-type Src potentiated c-Abl activation by growth factors, and a kinase-inactive form of Src reduced this activation, showing that Abl activation by growth factors occurs at least in part via activation of Src kinases. Significantly, we show that c-Abl has a functional role in the morphological response to PDGF. Whereas PDGF treatment of serum-starved wild-type mouse embryo fibroblasts resulted in distinct linear or circular/dorsal membrane ruffling, c-Abl-null cells demonstrated dramatically reduced ruffling in response to PDGF, which was rescued by physiological re-expression of c-Abl. These data identify c-Abl as a downstream target of activated receptor tyrosine kinases and Src family kinases, and show for the first time that c-Abl functions in the cellular response to growth factors.

Authors
Plattner, R; Kadlec, L; DeMali, KA; Kazlauskas, A; Pendergast, AM
MLA Citation
Plattner, R, Kadlec, L, DeMali, KA, Kazlauskas, A, and Pendergast, AM. "c-Abl is activated by growth factors and Src family kinases and has a role in the cellular response to PDGF." Genes Dev 13.18 (September 15, 1999): 2400-2411.
PMID
10500097
Source
pubmed
Published In
Genes & development
Volume
13
Issue
18
Publish Date
1999
Start Page
2400
End Page
2411

Efficient and rapid induction of a chronic myelogenous leukemia-like myeloproliferative disease in mice receiving P210 bcr/abl-transduced bone marrow.

Expression of the 210-kD bcr/abl fusion oncoprotein can cause a chronic myelogenous leukemia (CML)-like disease in mice receiving bone marrow cells transduced by bcr/abl-encoding retroviruses. However, previous methods failed to yield this disease at a frequency sufficient enough to allow for its use in the study of CML pathogenesis. To overcome this limitation, we have developed an efficient and reproducible method for inducing a CML-like disease in mice receiving P210 bcr/abl-transduced bone marrow cells. All mice receiving P210 bcr/abl-transduced bone marrow cells succumb to a myeloproliferative disease between 3 and 5 weeks after bone marrow transplantation. The myeloproliferative disease recapitulates many of the hallmarks of human CML and is characterized by high white blood cell counts and extensive extramedullary hematopoiesis in the spleen, liver, bone marrow, and lungs. Use of a retroviral vector coexpressing P210 bcr/abl and green fluorescent protein shows that the vast majority of bcr/abl-expressing cells are myeloid. Analysis of the proviral integration pattern shows that, in some mice, the myeloproliferative disease is clonal. In multiple mice, the CML-like disease has been transplantable, inducing a similar myeloproliferative syndrome within 1 month of transfer to sublethally irradiated syngeneic recipients. The disease in many of these mice has progressed to the development of acute lymphoma/leukemia resembling blast crisis. These results demonstrate that murine CML recapitulates important features of human CML. As such, it should be an excellent model for addressing specific issues relating to the pathogenesis and treatment of this disease.

Authors
Pear, WS; Miller, JP; Xu, L; Pui, JC; Soffer, B; Quackenbush, RC; Pendergast, AM; Bronson, R; Aster, JC; Scott, ML; Baltimore, D
MLA Citation
Pear, WS, Miller, JP, Xu, L, Pui, JC, Soffer, B, Quackenbush, RC, Pendergast, AM, Bronson, R, Aster, JC, Scott, ML, and Baltimore, D. "Efficient and rapid induction of a chronic myelogenous leukemia-like myeloproliferative disease in mice receiving P210 bcr/abl-transduced bone marrow." Blood 92.10 (November 15, 1998): 3780-3792.
PMID
9808572
Source
pubmed
Published In
Blood
Volume
92
Issue
10
Publish Date
1998
Start Page
3780
End Page
3792

Transforming properties of the P185, P210 and P230 variants of BCR/ABL in primary mouse bone marrow.

Authors
Quackenbush, RC; Reuther, GW; Pendergast, AM
MLA Citation
Quackenbush, RC, Reuther, GW, and Pendergast, AM. "Transforming properties of the P185, P210 and P230 variants of BCR/ABL in primary mouse bone marrow." November 15, 1998.
Source
wos-lite
Published In
Blood
Volume
92
Issue
10
Publish Date
1998
Start Page
217A
End Page
217A

Oncogenic Abl and Src tyrosine kinases elicit the ubiquitin-dependent degradation of target proteins through a Ras-independent pathway.

Oncogenic forms of the Abl and Src tyrosine kinases trigger the destruction of the Abi proteins, a family of Abl-interacting proteins that antagonize the oncogenic potential of Abl after overexpression in fibroblasts. The destruction of the Abi proteins requires tyrosine kinase activity and is dependent on the ubiquitin-proteasome pathway. We show that degradation of the Abi proteins occurs through a Ras-independent pathway. Significantly, expression of the Abi proteins is lost in cell lines and bone marrow cells isolated from patients with aggressive Bcr-Abl-positive leukemias. These findings suggest that loss of Abi proteins may be a component in the progression of Bcr-Abl-positive leukemias and identify a novel pathway linking activated nonreceptor protein tyrosine kinases to the destruction of specific target proteins through the ubiquitin-proteasome pathway.

Authors
Dai, Z; Quackenbush, RC; Courtney, KD; Grove, M; Cortez, D; Reuther, GW; Pendergast, AM
MLA Citation
Dai, Z, Quackenbush, RC, Courtney, KD, Grove, M, Cortez, D, Reuther, GW, and Pendergast, AM. "Oncogenic Abl and Src tyrosine kinases elicit the ubiquitin-dependent degradation of target proteins through a Ras-independent pathway." Genes Dev 12.10 (May 15, 1998): 1415-1424.
PMID
9585502
Source
pubmed
Published In
Genes & development
Volume
12
Issue
10
Publish Date
1998
Start Page
1415
End Page
1424

A requirement for NF-kappaB activation in Bcr-Abl-mediated transformation.

Bcr-Abl is a chimeric oncoprotein that is strongly implicated in acute lymphoblastic (ALL) and chronic myelogenous leukemias (CML). This deregulated tyrosine kinase selectively causes hematopoietic disorders resembling human leukemias in animal models and transforms fibroblasts and hematopoietic cells in culture. Bcr-Abl also protects cells from death induced on cytokine deprivation or exposure to DNA damaging agents. In addition, the antiapoptotic function of Bcr-Abl is thought to play a necessary role in hematopoietic transformation and potentially in leukemogenesis. The transcription factor NF-kappaB has been identified recently as an inhibitor of apoptosis and as a potential regulator of cellular transformation. This study shows that expression of Bcr-Abl leads to activation of NF-kappaB-dependent transcription by causing nuclear translocation of NF-kappaB as well as by increasing the transactivation function of the RelA/p65 subunit of NF-kappaB. Importantly, this activation is dependent on the tyrosine kinase activity of Bcr-Abl and partially requires Ras. The ability of Bcr-Abl to protect cytokine-dependent 32D myeloid cells from death induced by cytokine deprivation or DNA damage does not, however, require functional NF-kappaB. However, using a super-repressor form of IkappaBalpha, we show that NF-kappaB is required for Bcr-Abl-mediated tumorigenicity in nude mice and for transformation of primary bone marrow cells. This study implicates NF-kappaB as an important component of Bcr-Abl signaling. NF-kappaB-regulated genes, therefore, likely play a role in transformation by Bcr-Abl and thus in Bcr-Abl-associated human leukemias.

Authors
Reuther, JY; Reuther, GW; Cortez, D; Pendergast, AM; Baldwin, AS
MLA Citation
Reuther, JY, Reuther, GW, Cortez, D, Pendergast, AM, and Baldwin, AS. "A requirement for NF-kappaB activation in Bcr-Abl-mediated transformation." Genes Dev 12.7 (April 1, 1998): 968-981.
PMID
9531535
Source
pubmed
Published In
Genes & development
Volume
12
Issue
7
Publish Date
1998
Start Page
968
End Page
981

Protein tyrosine phosphatase 1B antagonizes signalling by oncoprotein tyrosine kinase p210 bcr-abl in vivo

The p210 bcr-abl protein tyrosine kinase (PTK) appears to be directly responsible for the initial manifestations of chronic myelogenous leukemia (CML). In contrast to the extensive characterization of the PTK and its effects on cell function, relatively little is known about the nature of the protein tyrosine phosphatases (PTPs) that may modulate p210 bcr-abl-induced signalling. In this study, we have demonstrated that expression of PTP1B is enhanced specifically in various cells expressing p210 bcr-abl, including a cell line derived from a patient with CML. This effect on expression of PTP1B required the kinase activity of p210 bcr-abl and occurred rapidly, concomitant with maximal activation of a temperature-sensitive mutant of the PTK. The effect is apparently specific for PTP1B since, among several PTPs tested, we detected no change in the levels of TCPTP, the closest relative of PTP1B. We have developed a strategy for identification of physiological substrates of individual PTPs which utilizes substrate-trapping mutant forms of the enzymes that retain the ability to bind to substrate but fail to catalyze efficient dephosphorylation. We have observed association between a substrate-trapping mutant of PTP1B (PTP1B-D181A) and p210 bcr-abl, but not v- Abl, in a cellular context. Consistent with the trapping data, we observed dephosphorylation of p210 bcr-abl, but not v-Abl, by PTP1B in vivo. We have demonstrated that PTP1B inhibited binding of the adapter protein Grb2 to p210 bcr-abl and suppressed p210 bcr-abl-induced transcriptional activation that is dependent on Ras. These results illustrate selectivity in the effects of PTPs in a cellular context and suggest that PTP1B may function as a specific, negative regulator of p210 bcr-abl signalling in vivo.

Authors
Jr, KRL; Flint, AJ; Jr, BRF; Pendergast, AM; Tonks, NK
MLA Citation
Jr, KRL, Flint, AJ, Jr, BRF, Pendergast, AM, and Tonks, NK. "Protein tyrosine phosphatase 1B antagonizes signalling by oncoprotein tyrosine kinase p210 bcr-abl in vivo." Molecular and Cellular Biology 18.5 (1998): 2965-2975.
PMID
9566916
Source
scival
Published In
Molecular and Cellular Biology
Volume
18
Issue
5
Publish Date
1998
Start Page
2965
End Page
2975

Cloning and characterization of the chronic neutrophilic leukemia-associated P230 BCR/ABL oncogene.

Authors
Quackenbush, RC; Reuther, GW; Pendergast, AM
MLA Citation
Quackenbush, RC, Reuther, GW, and Pendergast, AM. "Cloning and characterization of the chronic neutrophilic leukemia-associated P230 BCR/ABL oncogene." November 15, 1997.
Source
wos-lite
Published In
Blood
Volume
90
Issue
10
Publish Date
1997
Start Page
1087
End Page
1087

The amphiphysin-like protein 1 (ALP1) interacts functionally with the cABL tyrosine kinase and may play a role in cytoskeletal regulation.

cABL is a protooncogene, activated in a subset of human leukemias, whose protein product is a nonreceptor tyrosine kinase of unknown function. cABL has a complex structure that includes several domains and motifs found in proteins implicated in signal transduction pathways. An approach to elucidate cABL function is to identify proteins that interact directly with cABL and that may serve as regulators or effectors of its activity. To this end, a protein-interaction screen of a phage expression library was undertaken to identify proteins that interact with specific domains of cABL. An SH3-domain-containing protein has been identified that interacts with sequences in the cABL carboxyl terminus. The cDNA encoding ALP1 (amphiphysin-like protein 1) was isolated from a 16-day mouse embryo. ALP1 has high homology to BIN1, a recently cloned myc-interacting protein, and also shows significant homology to amphiphysin, a neuronal protein cloned from human and chicken. The amino terminus has homology to two yeast proteins, Rvs167 and Rvs161, which are involved in cell entry into stationary phase and cytoskeletal organization. ALP1 binds cABL in vitro and in vivo. Expression of ALP1 results in morphological transformation of NIH 3T3 fibroblasts in a cABL-dependent manner. The properties of ALP1 suggest that it may be involved in possible cytoskeletal functions of the cABL kinase. Additionally, these results provide further evidence for the importance of the cABL carboxyl terminus and its binding proteins in the regulation of cABL function.

Authors
Kadlec, L; Pendergast, AM
MLA Citation
Kadlec, L, and Pendergast, AM. "The amphiphysin-like protein 1 (ALP1) interacts functionally with the cABL tyrosine kinase and may play a role in cytoskeletal regulation." Proc Natl Acad Sci U S A 94.23 (November 11, 1997): 12390-12395.
PMID
9356459
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
94
Issue
23
Publish Date
1997
Start Page
12390
End Page
12395

The Bcr-Abl tyrosine kinase activates mitogenic signaling pathways and stimulates G1-to-S phase transition in hematopoietic cells.

Bcr-Abl is a constitutively active tyrosine kinase that is expressed in Philadelphia chromosome (Ph1)-positive human leukemias. Bcr-Abl has been shown to inhibit apoptosis and cause anchorage independent growth. However, its ability to activate mitogenic signaling pathways is controversial. Here we show that Bcr-Abl signaling prevents down-regulation of cyclin-dependent kinase activity and cell cycle arrest after growth factor deprivation of hematopoietic progenitor cells. Using an inducible system to regulate Bcr-Abl expression, we also demonstrate that Bcr-Abl expression is sufficient to induce G1-to-S phase transition, DNA synthesis, and activation of cyclin-dependent kinases in cells that were arrested in G0 by growth factor deprivation. Furthermore, Bcr-Abl activates Ras, Erk, and Jnk pathways as a primary consequence of expression. These data show that Bcr-Abl is one of a select group of oncogenes that is capable of both inhibiting apoptosis and deregulating cell proliferation. The combination of these activities is likely to be important for the progression of CML.

Authors
Cortez, D; Reuther, G; Pendergast, AM
MLA Citation
Cortez, D, Reuther, G, and Pendergast, AM. "The Bcr-Abl tyrosine kinase activates mitogenic signaling pathways and stimulates G1-to-S phase transition in hematopoietic cells." Oncogene 15.19 (November 6, 1997): 2333-2342.
PMID
9393877
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
15
Issue
19
Publish Date
1997
Start Page
2333
End Page
2342

The BCR-ABL tyrosine kinase inhibits apoptosis by activating a Ras-dependent signaling pathway.

BCR-ABL is a deregulated tyrosine kinase that is expressed in Philadelphia chromosome (Ph1) positive human leukemias. When expressed in hematopoietic cells, BCR-ABL causes cytokine independent proliferation, induces tumorigenic growth and prevents apoptosis in response to cytokine deprivation or DNA damage. One mechanism by which BCR-ABL signals in cells is by activating the small guanine nucleotide binding protein Ras. BCR-ABL-transformed cells have constitutively high levels of active, GTP-bound Ras. Here we use 32D cells that inducibly express a dominant negative Ras protein to define the Ras requirements in BCR-ABL-transformed cells. Dominant negative Ras inhibits BCR-ABL-mediated Ras activation, and induces cell death by an apoptotic mechanism. Therefore, BCR-ABL inhibits apoptosis through activation of a Ras-dependent signaling pathway.

Authors
Cortez, D; Stoica, G; Pierce, JH; Pendergast, AM
MLA Citation
Cortez, D, Stoica, G, Pierce, JH, and Pendergast, AM. "The BCR-ABL tyrosine kinase inhibits apoptosis by activating a Ras-dependent signaling pathway." Oncogene 13.12 (December 19, 1996): 2589-2594.
PMID
9000132
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
13
Issue
12
Publish Date
1996
Start Page
2589
End Page
2594

Functional association of CD7 with phosphatidylinositol 3-kinase: interaction via a YEDM motif.

Human CD7 is a 40 kDa protein expressed on thymocytes, early T, B, NK and myeloid lineage cells in bone marrow, and on mature T and NK cells. Previous studies suggested human CD7 may be involved in T and NK cell activation and/or adhesion, and that CD7-mediated cell activation may be transduced via the lipid kinase phosphatidylinositol 3-kinase (Pi3-kinase), a heterodimeric cytosolic protein consisting of an 85 kDa adaptor subunit that is coupled to a 110 kDa catalytic subunit. It has recently been shown that a sequence motif present in the cytoplasmic tall of both human and mouse CD7 bound with high affinity to recombinant SH2 domains present in the p85 subunit of Pi3-kinase. In this work, we used co-precipitation with anti-CD7 mAb 3A1 and recombinant p85 SH2-GST fusion proteins and peptide competition analysis to demonstrate that the cytoplasmic tail of CD7 interacts with a functional Pi3-kinase via the pTyr-X-X-Met motif. Furthermore, we show that cross-linking of CD7 markedly increased the amount of Pi3-kinase activity associated with CD7. The interaction of CD7 with the Pi3-kinase signal transduction pathway provides a mechanism for the previously observed functional responses attributed to CD7-mediated T and NK cell activation.

Authors
Lee, DM; Patel, DD; Pendergast, AM; Haynes, BF
MLA Citation
Lee, DM, Patel, DD, Pendergast, AM, and Haynes, BF. "Functional association of CD7 with phosphatidylinositol 3-kinase: interaction via a YEDM motif." Int Immunol 8.8 (August 1996): 1195-1203.
PMID
8918688
Source
pubmed
Published In
International Immunology
Volume
8
Issue
8
Publish Date
1996
Start Page
1195
End Page
1203

Nuclear tyrosine kinases: from Abl to WEE1.

The notion of a critical role for protein tyrosine kinases in the nucleus is supported by recent findings linking these proteins with components of the cell cycle and with the transcription machinery. Several of these tyrosine kinases localize to both nuclear and cytoplasmic compartments of the cell, and may coordinate signal transduction events in the cytoplasm with specific changes in the nucleus. Among these proteins are Abl, Rak, Fes and Fer. The past year has brought significant progress both towards the elucidation of the pathways that lead to activation of the Abl tyrosine kinase and towards the identification of novel Abl targets. Recent advances have also been made in understanding the regulation of the nucleus-specific human WEE1 tyrosine kinase. Nuclear tyrosine kinases may participate in the regulation of multiple cellular processes including transcription, DNA repair and the cell cycle.

Authors
Pendergast, AM
MLA Citation
Pendergast, AM. "Nuclear tyrosine kinases: from Abl to WEE1." Curr Opin Cell Biol 8.2 (April 1996): 174-181. (Review)
PMID
8791414
Source
pubmed
Published In
Current Opinion in Cell Biology
Volume
8
Issue
2
Publish Date
1996
Start Page
174
End Page
181

The roles of 14-3-3 proteins in signal transduction.

Authors
Reuther, GW; Pendergast, AM
MLA Citation
Reuther, GW, and Pendergast, AM. "The roles of 14-3-3 proteins in signal transduction." Vitam Horm 52 (1996): 149-175. (Review)
PMID
8909160
Source
pubmed
Published In
Vitamins and hormones
Volume
52
Publish Date
1996
Start Page
149
End Page
175

Dominant-negative mutants of Grb2 induced reversal of the transformed phenotypes caused by the point mutation-activated rat HER-2/Neu.

To clarify the role of the Shc-Grb2-Sos trimer in the oncogenic signaling of the point mutation-activated HER-2/neu receptor tyrosine kinase (named p185), we interfered with the protein-protein interactions in the Shc.Grb2.Sos complex by introducing Grb2 mutants with deletions in either amino- (delta N-Grb2) or carboxyl-(delta C-Grb2) terminal SH3 domains into B104-1-1 cells derived from NIH3T3 cells expressing the point mutation-activated HER-2/neu. We found that the transformed phenotypes of the B104-1-1 cells were largely reversed by the delta N-Grb2. The effect of the delta C-Grb2 was much weaker. Biochemical analysis showed that the delta N-Grb2 was able to associate Shc but not p185 or Sos, while the delta C-Grb2 bound to Shc, p185, and Sos. The p185-mediated Ras activation was severely inhibited by the delta N-Grb2 but not the delta C-Grb2. Taken together, these data demonstrate that interruption of the interaction between Shc and the endogenous Grb2 by the delta N-Grb2 impairs the oncogenic signaling of the activated p185, indicating that (i) the delta N-Grb2 functions as a strong dominant-negative mutant, and (ii) Shc/Grb2/Sos pathway plays a major role in mediating the oncogenic signal of the activated p185. Unlike the delta N-Grb2, delta C-Grb2 appears to be a relatively weak dominant-negative mutant, probably due to its ability to largely fulfill the biological functions of the wild-type Grb2.

Authors
Xie, Y; Pendergast, AM; Hung, MC
MLA Citation
Xie, Y, Pendergast, AM, and Hung, MC. "Dominant-negative mutants of Grb2 induced reversal of the transformed phenotypes caused by the point mutation-activated rat HER-2/Neu." J Biol Chem 270.51 (December 22, 1995): 30717-30724.
PMID
8530511
Source
pubmed
Published In
The Journal of biological chemistry
Volume
270
Issue
51
Publish Date
1995
Start Page
30717
End Page
30724

Mutant forms of growth factor-binding protein-2 reverse BCR-ABL-induced transformation.

Growth factor-binding protein 2 (Grb2) is an adaptor protein that links tyrosine kinases to Ras. BCR-ABL is a tyrosine kinase oncoprotein that is implicated in the pathogenesis of Philadelphia chromosome (Ph1)-positive leukemias. Grb2 forms a complex with BCR-ABL and the nucleotide exchange factor Sos that leads to the activation of the Ras protooncogene. In this report we demonstrate that Grb2 mutant proteins lacking amino- or carboxyl-terminal src homology SH3 domains suppress BCR-ABL-induced Ras activation and reverse the oncogenic phenotype. The Grb2 SH3-deletion mutant proteins bind to BCR-ABL and do not impair tyrosine kinase activity. Expression of the Grb2 SH3-deletion mutant proteins in BCR-ABL-transformed Rat-1 fibroblasts and in the human Ph1-positive leukemic cell line K562 inhibits their ability to grow as foci in soft agar and form tumors in nude mice. Furthermore, expression of the Grb2 SH3-deletion mutants in K562 cells induced their differentiation. Because Ras plays an important role in signaling by receptor and nonreceptor tyrosine kinases, the use of interfering mutant Grb2 proteins may be applied to block the proliferation of other cancers that depend in part on activated tyrosine kinases for growth.

Authors
Gishizky, ML; Cortez, D; Pendergast, AM
MLA Citation
Gishizky, ML, Cortez, D, and Pendergast, AM. "Mutant forms of growth factor-binding protein-2 reverse BCR-ABL-induced transformation." Proc Natl Acad Sci U S A 92.24 (November 21, 1995): 10889-10893.
PMID
7479904
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
92
Issue
24
Publish Date
1995
Start Page
10889
End Page
10893

Abi-2, a novel SH3-containing protein interacts with the c-Abl tyrosine kinase and modulates c-Abl transforming activity.

A protein has been identified that interacts specifically with both the Src homologous 3 (SH3) domain and carboxy-terminal sequences of the c-Abl tyrosine kinase. The cDNA encoding the Abl interactor protein (Abi-2), was isolated from a human lymphocyte library using the yeast two-hybrid system with the Abl SH3 domain as bait. Abi-2 binds to c-Abl in vitro and in vivo. Abi-2 is a novel protein that contains an SH3 domain and proline-rich sequences critical for binding to c-Abl. A basic region in the amino terminus of Abi-2 is homologous to the DNA-binding sequence of homeo-domain proteins. We show that Abi-2 is a substrate for the c-Abl tyrosine kinase. Expression of an Abi-2 mutant protein that lacks sequences required for binding to the Abl SH3 domain but retains binding to the Abl carboxyl terminus activates the transforming capacity of c-Abl. The properties of Abi-2 are consistent with a dual role as regulator and potential effector of the c-Abl protein and suggest that Abi-2 may function as a tumor suppressor in mammalian cells.

Authors
Dai, Z; Pendergast, AM
MLA Citation
Dai, Z, and Pendergast, AM. "Abi-2, a novel SH3-containing protein interacts with the c-Abl tyrosine kinase and modulates c-Abl transforming activity." Genes Dev 9.21 (November 1, 1995): 2569-2582.
PMID
7590236
Source
pubmed
Published In
Genes & development
Volume
9
Issue
21
Publish Date
1995
Start Page
2569
End Page
2582

Structural and signaling requirements for BCR-ABL-mediated transformation and inhibition of apoptosis.

BCR-ABL is a deregulated tyrosine kinase expressed in Philadelphia chromosome-positive human leukemias. Prolongation of hematopoietic cell survival by inhibition of apoptosis has been proposed to be an integral component of BCR-ABL-induced chronic myelogenous leukemia. BCR-ABL elicits transformation of both fibroblast and hematopoietic cells and blocks apoptosis following cytokine deprivation in various factor-dependent cells. To elucidate the mechanisms whereby BCR-ABL induces transformation and blocks apoptosis in hematopoietic cells, we examined the biological effects of expression of a series of BCR-ABL mutants. Single amino acid substitutions in the GRB2 binding site (Y177F), Src homology 2 domain (R552L), or an autophosphorylation site in the tyrosine kinase domain (Y793F) do not diminish the antiapoptotic and transforming properties of BCR-ABL in hematopoietic cells, although these mutations were previously shown to drastically reduce the transforming activity of BCR-ABL in fibroblasts. A BCR-ABL molecule containing all three mutations (Y177F/R552L/Y793F) exhibits a severe decrease in transforming and antiapoptotic activities compared with the wild-type BCR-ABL protein in 32D myeloid progenitor cells. Ras is activated, the SHC adapter protein is tyrosine phosphorylated and binds GRB2, and myc mRNA levels are increased following expression of all kinase active BCR-ABL proteins with the exception of the Y177F/R552L/Y793F BCR-ABL mutant in 32D cells. We propose that BCR-ABL uses multiple pathways to activate Ras in hematopoietic cells and that this activation is necessary for the transforming and antiapoptotic activities of BCR-ABL. However, Ras activation is not sufficient for BCR-ABL-mediated transformation. A BCR-ABL deletion mutant (delta 176-427) that activates Ras and blocks apoptosis but has severely impaired transforming ability in 32D cells has been identified. These data suggest that BCR-ABL requires additional signaling components to elicit tumorigenic growth which are distinct from those required to block apoptosis.

Authors
Cortez, D; Kadlec, L; Pendergast, AM
MLA Citation
Cortez, D, Kadlec, L, and Pendergast, AM. "Structural and signaling requirements for BCR-ABL-mediated transformation and inhibition of apoptosis." Mol Cell Biol 15.10 (October 1995): 5531-5541.
PMID
7565705
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
15
Issue
10
Publish Date
1995
Start Page
5531
End Page
5541

ENHANCED EXPRESSION OF A PROTEIN-TYROSINE-PHOSPHATASE IN CELLS EXPRESSING P210BCR/ABL

Authors
LAMONTAGNE, KR; PENDERGAST, AM; FRANZA, BR; TONKS, NK
MLA Citation
LAMONTAGNE, KR, PENDERGAST, AM, FRANZA, BR, and TONKS, NK. "ENHANCED EXPRESSION OF A PROTEIN-TYROSINE-PHOSPHATASE IN CELLS EXPRESSING P210BCR/ABL." January 5, 1995.
Source
wos-lite
Published In
Journal of Cellular Biochemistry
Publish Date
1995
Start Page
65
End Page
65

Abi-2, a novel SH3-containing protein interacts with the c-Abl tyrosine kinase and modulates c-Abl transforming activity

A protein has been identified that interacts specifically with both the Src homologous 3 (SH3) domain and carboxy-terminal sequences of the c-Abl tyrosine kinase. The cDNA encoding the Abl interactor protein (Abi-2), was isolated from a human lymphocyte library using the yeast two-hybrid system with the Abl SH3 domain as bait. Abi-2 binds to c-Abl in vitro and in vivo. Abi-2 is a novel protein that contains an SH3 domain and proline-rich sequences critical for binding to c-Abl. A basic region in the amino terminus of Abi-2 is homologous to the DNA-binding sequence of homeo-domain proteins. We show that Abi-2 is a substrate for the c-Abl tyrosine kinase. Expression of an Abi-2 mutant protein that lacks sequences required for binding to the Abl SH3 domain but retains binding to the Abl carboxyl terminus activates the transforming capacity of c-Abl. The properties of Abi-2 are consistent with a dual role as regulator and potential effector of the c-Abl protein and suggest that Abi-2 may function as a tumor suppressor in mammalian cells.

Authors
Dai, Z; Pendergast, AM
MLA Citation
Dai, Z, and Pendergast, AM. "Abi-2, a novel SH3-containing protein interacts with the c-Abl tyrosine kinase and modulates c-Abl transforming activity." Genes and Development 9.21 (1995): X-2582.
Source
scival
Published In
Genes and Development
Volume
9
Issue
21
Publish Date
1995
Start Page
X
End Page
2582

Association of the protein kinases c-Bcr and Bcr-Abl with proteins of the 14-3-3 family.

In this study, a protein that interacts with sequences encoded by the first exon of the protein kinase Bcr was cloned. The Bcr-associated protein 1 (Bap-1) is a member of the 14-3-3 family of proteins. Bap-1 interacts with full-length c-Bcr and with the chimeric Bcr-Abl tyrosine kinase of Philadelphia chromosome (Ph1)-positive human leukemias. Bap-1 is a substrate for the Bcr serine-threonine kinase and is also phosphorylated on tyrosine by Bcr-Abl but not by c-Abl. Bap-1 may function in the regulation of c-Bcr and may contribute to the transforming activity of Bcr-Abl in vivo. 14-3-3 proteins are essential for cell proliferation and have a role in determining the timing of mitosis in yeast. Through direct binding to sequences present in Bcr and in other proteins implicated in signaling, the mammalian 14-3-3 proteins may link specific signaling protein components to mitogenic and cell-cycle control pathways.

Authors
Reuther, GW; Fu, H; Cripe, LD; Collier, RJ; Pendergast, AM
MLA Citation
Reuther, GW, Fu, H, Cripe, LD, Collier, RJ, and Pendergast, AM. "Association of the protein kinases c-Bcr and Bcr-Abl with proteins of the 14-3-3 family." Science 266.5182 (October 7, 1994): 129-133.
PMID
7939633
Source
pubmed
Published In
Science
Volume
266
Issue
5182
Publish Date
1994
Start Page
129
End Page
133

BCR-ABL-induced oncogenesis is mediated by direct interaction with the SH2 domain of the GRB-2 adaptor protein.

BCR-ABL is a chimeric oncoprotein that exhibits deregulated tyrosine kinase activity and is implicated in the pathogenesis of Philadelphia chromosome (Ph1)-positive human leukemias. Sequences within the first exon of BCR are required to activate the transforming potential of BCR-ABL. The SH2/SH3 domain-containing GRB-2 protein links tyrosine kinases to Ras signaling. We demonstrate that BCR-ABL exists in a complex with GRB-2 in vivo. Binding of GRB-2 to BCR-ABL is mediated by the direct interaction of the GRB-2 SH2 domain with a phosphorylated tyrosine, Y177, within the BCR first exon. The BCR-ABL-GRB-2 interaction is required for activation of the Ras signaling pathway. Mutation of Y177 to phenylalanine (Y177F) abolishes GRB-2 binding and abrogates BCR-ABL-induced Ras activation. The BCR-ABL (Y177F) mutant is unable to transform primary bone marrow cultures and is impaired in its ability to transform Rat1 fibroblasts. These findings implicate activation of Ras function as an important component in BCR-ABL-mediated transformation and demonstrate that GRB-2 not only functions in normal development and mitogenesis but also plays a role in oncogenesis.

Authors
Pendergast, AM; Quilliam, LA; Cripe, LD; Bassing, CH; Dai, Z; Li, N; Batzer, A; Rabun, KM; Der, CJ; Schlessinger, J
MLA Citation
Pendergast, AM, Quilliam, LA, Cripe, LD, Bassing, CH, Dai, Z, Li, N, Batzer, A, Rabun, KM, Der, CJ, and Schlessinger, J. "BCR-ABL-induced oncogenesis is mediated by direct interaction with the SH2 domain of the GRB-2 adaptor protein." Cell 75.1 (October 8, 1993): 175-185.
PMID
8402896
Source
pubmed
Published In
Cell
Volume
75
Issue
1
Publish Date
1993
Start Page
175
End Page
185

Oncogenic activation of c-ABL by mutation within its last exon.

The c-ABL proto-oncogene is a predominantly nuclear localized tyrosine kinase. A random mutagenesis scheme was used to isolate c-ABL mutants whose expression produced a transformed phenotype in rodent fibroblast cells. An in-frame deletion within the central region of the last exon was identified in one ABL mutant. The mechanism of c-ABL oncogenic activation by mutation within the last exon differs both functionally and structurally from those of v-ABL and BCR/ABL. This class of ABL mutants shows increased tyrosine phosphorylation of cellular proteins in vivo but low levels of autophosphorylation. Last-exon ABL mutants are distinguished from v-ABL or BCR/ABL by their inability to transform primary bone marrow cells or support the growth of transformed pre-B cells. These findings define a new mechanism of oncogenic activation for the ABL kinase through mutations in the last exon which do not require amino-terminal deletions or mutations within the src homology regions.

Authors
Goga, A; McLaughlin, J; Pendergast, AM; Parmar, K; Muller, A; Rosenberg, N; Witte, ON
MLA Citation
Goga, A, McLaughlin, J, Pendergast, AM, Parmar, K, Muller, A, Rosenberg, N, and Witte, ON. "Oncogenic activation of c-ABL by mutation within its last exon." Mol Cell Biol 13.8 (August 1993): 4967-4975.
PMID
8336729
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
13
Issue
8
Publish Date
1993
Start Page
4967
End Page
4975

En bloc substitution of the Src homology region 2 domain activates the transforming potential of the c-Abl protein tyrosine kinase.

Src homology region 2 (SH2) domains are present in many proteins involved in signal transduction. In nonreceptor protein tyrosine kinases the SH2 domain has been implicated in regulation of tyrosine kinase activity and in mediating interactions involved in downstream signaling. Different SH2 domains exhibit distinct binding specificities for both phosphotyrosine- and phosphoserine/phosphothreonine-containing proteins. We show that different SH2 domains are not functionally equivalent within the context of the c-ABL1b protooncogene. c-ABL1b, altered by replacement of its SH2 domain with the N-terminal SH2 domain of Ras GTPase-activating protein, exhibited activated transforming capability, caused intracellular tyrosine phosphorylation of p62, and was relocalized from nucleus to cytoplasm. This en bloc substitution apparently uncouples two distinct functions of the SH2 domain so that c-ABL escapes normal regulatory control while it retains the capability to elicit signals that promote transformation. The SH2 domain of the ARG protein tyrosine kinase, which shares high amino acid-sequence homology with the SH2 domain of ABL, was less effective in activating the oncogenic potential of c-ABL. The effects that the N-terminal SH2 domain of Ras GTPase-activating protein has in the context of c-ABL resemble the effects of deleting the SH3 domain. Thus, the SH2 and SH3 domains may have coordinate roles as regulatory control elements within the context of c-ABL.

Authors
Muller, AJ; Pendergast, AM; Parmar, K; Havlik, MH; Rosenberg, N; Witte, ON
MLA Citation
Muller, AJ, Pendergast, AM, Parmar, K, Havlik, MH, Rosenberg, N, and Witte, ON. "En bloc substitution of the Src homology region 2 domain activates the transforming potential of the c-Abl protein tyrosine kinase." Proc Natl Acad Sci U S A 90.8 (April 15, 1993): 3457-3461.
PMID
7682703
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
90
Issue
8
Publish Date
1993
Start Page
3457
End Page
3461

SH1 domain autophosphorylation of P210 BCR/ABL is required for transformation but not growth factor independence.

P210 BCR/ABL is a chimeric oncogene implicated in the pathogenesis of chronic myelogenous leukemia. BCR sequences have been shown to be required for activation of the tyrosine kinase and transforming functions of BCR/ABL. In this work, we show that two other structural requirements for full transforming activity of P210 BCR/ABL include a functional tyrosine kinase and the presence of tyrosine 1294, a site of autophosphorylation within the tyrosine kinase domain. Replacement of tyrosine 1294 with phenylalanine (1294F) greatly diminishes the transforming activity of BCR/ABL without affecting the specific activity of the protein tyrosine kinase. Expression of an exogenous myc gene in fibroblasts partially complements the transforming capacity of mutant P210 BCR/ABL (1294F). Surprisingly, tyrosine 1294 is not required for efficient induction of growth factor-independence in hematopoietic cell lines by P210 BCR/ABL. These results suggest that autophosphorylation at tyrosine 1294 may be important for recognition and phosphorylation of cellular substrates in the pathway of transformation, but it is not critical for mediating the events which lead to growth factor independence.

Authors
Pendergast, AM; Gishizky, ML; Havlik, MH; Witte, ON
MLA Citation
Pendergast, AM, Gishizky, ML, Havlik, MH, and Witte, ON. "SH1 domain autophosphorylation of P210 BCR/ABL is required for transformation but not growth factor independence." Mol Cell Biol 13.3 (March 1993): 1728-1736.
PMID
8441409
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
13
Issue
3
Publish Date
1993
Start Page
1728
End Page
1736

A limited set of SH2 domains binds BCR through a high-affinity phosphotyrosine-independent interaction.

SH2 (src homology region 2) domains are implicated in protein-protein interactions involved in signal transduction pathways. Isolated SH2 domains bind proteins that are tyrosine phosphorylated. A novel, phosphotyrosine-independent binding interaction between BCR, the Philadelphia chromosome breakpoint cluster region gene product, and the SH2 domain of its translocation partner c-ABL has recently been reported. We have examined the ability of additional SH2 domains to bind phosphotyrosine-free BCR and compared this with their ability to bind tyrosine-phosphorylated c-ABL 1b. Of 11 individual SH2 domains examined, 8 exhibited relatively high affinity for c-ABL 1b, whereas only 4 exhibited relatively high affinity for BCR. Binding of tyrosine-phosphorylated c-ABL 1b by the relatively high-affinity ABL and ARG SH2 domains was quantitatively analyzed, and equilibrium dissociation constants for both interactions were estimated to be in the range of 5 x 10(-7) M. The ABL SH2 domain exhibited relatively high affinity for phosphotyrosine-free BCR as well; however, this interaction appears to be about two orders of magnitude weaker than binding of tyrosine-phosphorylated c-ABL 1b. The ARG SH2 domain exhibited relatively weak affinity for BCR and was determined to bind about 10-fold less strongly than the ABL SH2 domain. The ABL and ARG SH2 domains differ by only 10 of 91 amino acids, and the substitution of ABL-specific amino acids into either the amino- or carboxy-terminal half of the ARG SH2 domain was found to increase its affinity for BCR. We discuss these results in terms of a model which has been proposed for peptide binding by class I histocompatibility glycoproteins.

Authors
Muller, AJ; Pendergast, AM; Havlik, MH; Puil, L; Pawson, T; Witte, ON
MLA Citation
Muller, AJ, Pendergast, AM, Havlik, MH, Puil, L, Pawson, T, and Witte, ON. "A limited set of SH2 domains binds BCR through a high-affinity phosphotyrosine-independent interaction." Mol Cell Biol 12.11 (November 1992): 5087-5093.
PMID
1383690
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
12
Issue
11
Publish Date
1992
Start Page
5087
End Page
5093

The 5' non-coding region of the BCR/ABL oncogene augments its ability to stimulate the growth of immature lymphoid cells.

The Philadelphia chromosome (Ph1, t9:22;34:q11) is a reciprocal translocation between chromosome 22 and chromosome 9 which results in the formation of the chimeric BCR/ABL oncogene. Alternative forms of BCR/ABL are produced by splicing different sets of exons of the BCR gene to a common set of c-ABL sequences. This results in the formation of an 8.7 kilobase mRNA that encodes the P210 BCR/ABL gene product or a 7.0 kilobase mRNA that encodes the P185 BCR/ABL gene product. Both BCR/ABL transcripts derive their 5' non-coding sequences from the BCR gene locus. This 5' region is over 500 nucleotides in length, has a GC content greater than 75% and has a short open reading frame. To determine if this unusual 5' non-coding region plays a role in BCR/ABL transformation, we prepared retroviral vectors containing identical BCR/ABL coding regions but differing in the length of the BCR 5' non-coding region. Matched viral stocks were evaluated for their ability to transform bone marrow in vitro and for their ability to cause tumors when inoculated into 3- to 4-week-old mice. In this report we present the unexpected finding that the BCR/ABL 5' non-coding region augments the transforming activity of both P210 and P185 BCR/ABL in vitro. In vivo, BCR/ABL is a weak tumorigenic agent and its potency is enhanced by the presence of the 5' non-coding region.

Authors
Gishizky, ML; McLaughlin, J; Pendergast, AM; Witte, ON
MLA Citation
Gishizky, ML, McLaughlin, J, Pendergast, AM, and Witte, ON. "The 5' non-coding region of the BCR/ABL oncogene augments its ability to stimulate the growth of immature lymphoid cells." Oncogene 6.8 (August 1991): 1299-1306.
PMID
1886706
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
6
Issue
8
Publish Date
1991
Start Page
1299
End Page
1306

BCR sequences essential for transformation by the BCR-ABL oncogene bind to the ABL SH2 regulatory domain in a non-phosphotyrosine-dependent manner.

BCR-ABL is a chimeric oncogene implicated in the pathogenesis of Philadelphia chromosome-positive human leukemias. BCR first exon sequences specifically activate the tyrosine kinase and transforming potential of BCR-ABL. We have tested the hypothesis that activation of BCR-ABL may involve direct interaction between BCR sequences and the tyrosine kinase regulatory domains of ABL. Full-length c-BCR as well as BCR sequences retained in BCR-ABL bind specifically to the SH2 domain of ABL. The binding domain has been localized within the first exon of BCR and consists of at least two SH2-binding sites. This domain is essential for BCR-ABL-mediated transformation. Phosphoserine/phosphothreonine but not phosphotyrosine residues on BCR are required for interaction with the ABL SH2 domain. These findings extend the range of potential SH2-protein interactions in growth control pathways and suggest a function for SH2 domains in the activation of the BCR-ABL oncogene as well as a role for BCR in cellular signaling pathways.

Authors
Pendergast, AM; Muller, AJ; Havlik, MH; Maru, Y; Witte, ON
MLA Citation
Pendergast, AM, Muller, AJ, Havlik, MH, Maru, Y, and Witte, ON. "BCR sequences essential for transformation by the BCR-ABL oncogene bind to the ABL SH2 regulatory domain in a non-phosphotyrosine-dependent manner." Cell 66.1 (July 12, 1991): 161-171.
PMID
1712671
Source
pubmed
Published In
Cell
Volume
66
Issue
1
Publish Date
1991
Start Page
161
End Page
171

Evidence for regulation of the human ABL tyrosine kinase by a cellular inhibitor.

Phosphotyrosine cannot be detected on normal human ABL protein-tyrosine kinases, but activated oncogenic forms of the human ABL protein are phosphorylated on tyrosine in vivo. Activation of ABL can occur by substitution of the ABL first exon with breakpoint cluster region (BCR) sequences or by deletion of the noncatalytic SH3 (src homology region 3) domain. An alternative mode for the activation of the ABL kinases is hyperexpression at greater than 500-fold over endogenous levels. This is not a consequence of transphosphorylation of the hyperexpressed ABL molecules. ABL proteins translated in vitro lack phosphotyrosine, but tyrosine kinase activity is uncovered after immunoprecipitation and removal of lysate components. The rates of dephosphorylation of ABL and BCR-ABL fusion protein by phosphotyrosine-specific phosphatases are approximately the same. These combined results indicate that inhibition of ABL activity is reversible and suggest that a cellular component interacts noncovalently with ABL to inhibit its autophosphorylation.

Authors
Pendergast, AM; Muller, AJ; Havlik, MH; Clark, R; McCormick, F; Witte, ON
MLA Citation
Pendergast, AM, Muller, AJ, Havlik, MH, Clark, R, McCormick, F, and Witte, ON. "Evidence for regulation of the human ABL tyrosine kinase by a cellular inhibitor." Proc Natl Acad Sci U S A 88.13 (July 1, 1991): 5927-5931.
PMID
1712111
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
88
Issue
13
Publish Date
1991
Start Page
5927
End Page
5931

BCR first exon sequences specifically activate the BCR/ABL tyrosine kinase oncogene of Philadelphia chromosome-positive human leukemias.

The c-abl proto-oncogene encodes a cytoplasmic tyrosine kinase which is homologous to the src gene product in its kinase domain and in the upstream kinase regulatory domains SH2 (src homology region 2) and SH3 (src homology region 3). The murine v-abl oncogene product has lost the SH3 domain as a consequence of N-terminal fusion of gag sequences. Deletion of the SH3 domain is sufficient to render the murine c-abl proto-oncogene product transforming when myristylated N-terminal membrane localization sequences are also present. In contrast, the human BCR/ABL oncogene of the Philadelphia chromosome translocation has an intact SH3 domain and its product is not myristylated at the N terminus. To analyze the contribution of BCR-encoded sequences to BCR/ABL-mediated transformation, the effects of a series of deletions and substitutions were assessed in fibroblast and hematopoietic-cell transformation assays. BCR first-exon sequences specifically potentiate transformation and tyrosine kinase activation when they are fused to the second exon of otherwise intact c-ABL. This suggests that BCR-encoded sequences specifically interfere with negative regulation of the ABL-encoded tyrosine kinase, which would represent a novel mechanism for the activation of nonreceptor tyrosine kinase-encoding proto-oncogenes.

Authors
Muller, AJ; Young, JC; Pendergast, AM; Pondel, M; Landau, NR; Littman, DR; Witte, ON
MLA Citation
Muller, AJ, Young, JC, Pendergast, AM, Pondel, M, Landau, NR, Littman, DR, and Witte, ON. "BCR first exon sequences specifically activate the BCR/ABL tyrosine kinase oncogene of Philadelphia chromosome-positive human leukemias." Mol Cell Biol 11.4 (April 1991): 1785-1792.
PMID
2005881
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
11
Issue
4
Publish Date
1991
Start Page
1785
End Page
1792

Tyrosine kinase activity and transformation potency of bcr-abl oncogene products.

Oncogenic activation of the proto-oncogene c-abl in human leukemias occurs as a result of the addition of exons from the gene bcr and truncation of the first abl exon. Analysis of tyrosine kinase activity and quantitative measurement of transformation potency in a single-step assay indicate that variation in bcr exon contribution results in a functional difference between p210bcr-abl and p185bcr-abl proteins. Thus, foreign upstream sequences are important in the deregulation of the kinase activity of the abl product, and the extent of deregulation correlates with the pathological effects of the bcr-abl proteins.

Authors
Lugo, TG; Pendergast, AM; Muller, AJ; Witte, ON
MLA Citation
Lugo, TG, Pendergast, AM, Muller, AJ, and Witte, ON. "Tyrosine kinase activity and transformation potency of bcr-abl oncogene products." Science 247.4946 (March 2, 1990): 1079-1082.
PMID
2408149
Source
pubmed
Published In
Science
Volume
247
Issue
4946
Publish Date
1990
Start Page
1079
End Page
1082

P150c-abl is detected in mouse male germ cells by an in vitro kinase assay and is associated with stage-specific phosphoproteins in haploid cells.

We have utilized c-abl antibodies and an in vitro kinase assay to identify the protein products of the c-abl gene in mouse testis. Although the testis contains high levels of a unique c-abl mRNA, along with lower amounts of two c-abl mRNAs common to somatic cells, we detected only a single polypeptide of approximately 150 kd, indistinguishable by our methods from P150c-abl observed in murine tissues and cell lines. P150c-abl was also detected in enriched populations of germ cells, including late stage spermatids which contain the highest levels of the novel c-abl transcript. In mature testis, and specifically in late spermatids, P150c-abl co-precipitated with phosphoproteins of approximately 74 kd which were labeled during the in vitro kinase assay. These proteins were phosphorylated predominantly on serine and their phosphopeptide maps differed from that of P150c-abl. The P74 phosphoproteins were not found in association with P150c-abl in germ cells at earlier developmental stages, nor in other tissues or cell lines examined.

Authors
Ponzetto, C; Wadewitz, AG; Pendergast, AM; Witte, ON; Wolgemuth, DJ
MLA Citation
Ponzetto, C, Wadewitz, AG, Pendergast, AM, Witte, ON, and Wolgemuth, DJ. "P150c-abl is detected in mouse male germ cells by an in vitro kinase assay and is associated with stage-specific phosphoproteins in haploid cells." Oncogene 4.6 (June 1989): 685-690.
PMID
2660070
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
4
Issue
6
Publish Date
1989
Start Page
685
End Page
690

Baculovirus expression of functional P210 BCR-ABL oncogene product.

The chronic myelogenous leukemia-associated P210 BCR-ABL oncogene protein product has been produced using the baculovirus expression system. High-level expression of the P210 BCR-ABL protein required the removal of GC rich 5' non-coding sequences. P210 BCR-ABL synthesized in insect cells is an active tyrosine protein kinase indistinguishable from P210 BCR-ABL isolated from human cells. Both proteins utilize angiotensin II as a phosphate acceptor in vitro with a Km for ATP of approximately 1.5 microM. P210 BCR-ABL produced in insect cells undergoes autophosphorylation in vitro and in vivo. Gel filtration of P210 BCR-ABL reveals that the protein elutes as a high molecular weight complex of about 800 kD. Approximately 4 to 5 mg of P210 BCR-ABL is produced in one liter of infected insect cells. Following cell disruption and a three-step ion exchange and gel filtration purification procedure, 0.4 mg of soluble P210 BCR-ABL is obtained per liter of suspension culture. An alternative procedure employing detergent extraction and immunoaffinity chromatography gave higher yields and purity from smaller amounts of infected cell extracts. The availability of intact, soluble and enzymatically active P210 BCR-ABL represents a significant advance for studying the biochemical and biophysical properties of the ABL oncogene family of proteins.

Authors
Pendergast, AM; Clark, R; Kawasaki, ES; McCormick, FP; Witte, ON
MLA Citation
Pendergast, AM, Clark, R, Kawasaki, ES, McCormick, FP, and Witte, ON. "Baculovirus expression of functional P210 BCR-ABL oncogene product." Oncogene 4.6 (June 1989): 759-766.
PMID
2499863
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
4
Issue
6
Publish Date
1989
Start Page
759
End Page
766

Expression of a distinctive BCR-ABL oncogene in Ph1-positive acute lymphocytic leukemia (ALL).

The Philadelphia chromosome (Ph1) is a translocation between chromosomes 9 and 22 that is found in chronic myelogenous leukemia (CML) and a subset of acute lymphocytic leukemia patients (ALL). In CML, this results in the expression of a chimeric 8.5-kilobase BCR-ABL transcript that encodes the P210BCR-ABL tyrosine kinase. The Ph1 chromosome in ALL expresses a distinct ABL-derived 7-kilobase messenger RNA that encodes the P185ALL-ABL protein. Since the expression of different oncogene products may play a role in the distinctive presentation of Ph1-positive ALL versus CML, it is necessary to understand the molecular basis for the expression of P185ALL-ABL. Both P210BCR-ABL and P185ALL-ABL are recognized by an antiserum directed to BCR determinants in the amino-terminal region of both proteins. Antisera to BCR determinants proximal to the BCR-ABL junction in CML immunoprecipitated P210BCR-ABL but not P185ALL-ABL. Nucleotide sequence analysis of complementary DNA clones made from RNA from the Ph1-positive ALL SUP-B15 cell line, and S1 nuclease protection analysis confirmed the presence of BCR-ABL chimeric transcripts in Ph1-positive ALL cells. In Ph1-positive ALL, ABL sequences were joined to BCR sequences approximately 1.5 kilobases 5' of the CML junction. P185ALL-ABL represents the product of a BCR-ABL fusion gene in Ph1-positive ALL that is distinct from the BCR-ABL fusion gene of CML.

Authors
Clark, SS; McLaughlin, J; Timmons, M; Pendergast, AM; Ben-Neriah, Y; Dow, LW; Crist, W; Rovera, G; Smith, SD; Witte, ON
MLA Citation
Clark, SS, McLaughlin, J, Timmons, M, Pendergast, AM, Ben-Neriah, Y, Dow, LW, Crist, W, Rovera, G, Smith, SD, and Witte, ON. "Expression of a distinctive BCR-ABL oncogene in Ph1-positive acute lymphocytic leukemia (ALL)." Science 239.4841 Pt 1 (February 12, 1988): 775-777.
PMID
3422516
Source
pubmed
Published In
Science
Volume
239
Issue
4841 Pt 1
Publish Date
1988
Start Page
775
End Page
777

Expression of a distinctive BCR-ABL oncogene in Ph1-positive acute lymphocytic leukemia (ALL)

The Philadelphia chromosome (Ph1) is a translocation between chromosomes 9 and 22 that is found in chronic myelogenous leukemia (CML) and a subset of acute lymphocytic leukemia patients (ALL). In CML, this results in the expression of a chimeric 8.5-kilobase BCR-ABL transcript that encodes the P210(BCR-ABL) tyrosine kinase. The Ph1 chromosome in ALL expresses a distinct ABL-derived 7-kilobase messenger RNA that encodes the P185(ALL-ABL) protein. Since the expression of different oncogene products may play a role in the distinctive presentation of Ph1-positive ALL versus CML, it is necessary to understand the molecular basis for the expression of P185(ALL-ABL). Both P210(BCR-ABL) and P185(ALL-ABL) are recognized by an antiserum directed to BCR determinants in the amino-terminal region of both proteins. Antisera to BCR determinants proximal to the BCR-ABL junction in CML immunoprecipitated P210(BCR-ABL) but not P185(ALL-ABL). Nucleotide sequence analysis of complementary DNA clones made from RNA from the Ph1-positive ALL SUP-B15 cell line, and S1 nuclease protection analysis confirmed the presence of BCR-ABL chimeric transcripts in PH1-positive ALL cells. In Ph1-positive ALL, ABL sequences were joined to BCR sequences approximately 1.5 kilobases 5' of the CML junction. P185(ALL-ABL) represents the product of a BCR-ABL fusion gene in Ph1-positive ALL that is distinct from the BCR-ABL fusion gene of CML.

Authors
Clark, SS; McLaughlin, J; Timmons, M; Pendergast, AM; Ben-Neriah, Y; Dow, LW; Crist, W; Rovera, G; Smith, SD; Witte, ON
MLA Citation
Clark, SS, McLaughlin, J, Timmons, M, Pendergast, AM, Ben-Neriah, Y, Dow, LW, Crist, W, Rovera, G, Smith, SD, and Witte, ON. "Expression of a distinctive BCR-ABL oncogene in Ph1-positive acute lymphocytic leukemia (ALL)." Science 239.4841 (1988): 775-777.
Source
scival
Published In
Science
Volume
239
Issue
4841
Publish Date
1988
Start Page
775
End Page
777

Role of the ABL oncogene tyrosine kinase activity in human leukaemia.

A great deal of information has emerged over the past decade regarding the gene structures and corresponding protein products of the cellular and transformation-associated forms of the ABL tyrosine kinase family. Many reports have also detailed the biological effects of these proteins (particularly the viral ABL forms) on a broad range of cell types. However, in spite of all these research efforts, the precise role of the ABL gene in normal and neoplastic growth remains to be determined. To elucidate the mechanism of action of normal and altered ABL proteins, it is imperative to identify their relevant cellular substrates and establish the role of the ABL target proteins in transformation and normal cellular growth. The availability of temperature-sensitive ABL proteins, coupled with the use of sensitive anti-phosphotyrosine antibodies, should be useful in this respect. Purification of enzymatically active, intact forms of the ABL proteins produced in insect cells by employing baculovirus expression vectors should permit direct comparison of the biochemical properties and tertiary structures of the various members of the ABL protein kinase family. Such studies will aid in understanding the nature of the alteration of ABL which results in the activation of its transforming potential. Furthermore, the availability of purified ABL proteins should permit examination of interactions of ABL with other growth-regulatory proteins, such as growth factor receptors. It has been shown that transformation-associated ABL proteins interact with the IL-3, IL-2 and GM-CSF growth-factor pathways. These and other components of the cellular signalling pathways are potential ABL targets. The elucidation of ABL function by a variety of approaches such as those described above will ultimately aid in the development of far-reaching therapeutic treatments for at least two forms of human leukaemia: Ph positive CML and Ph positive ALL.

Authors
Pendergast, AM; Witte, ON
MLA Citation
Pendergast, AM, and Witte, ON. "Role of the ABL oncogene tyrosine kinase activity in human leukaemia." Baillieres Clin Haematol 1.4 (December 1987): 1001-1020. (Review)
PMID
3332851
Source
pubmed
Published In
Best Practice and Research: Clinical Haematology
Volume
1
Issue
4
Publish Date
1987
Start Page
1001
End Page
1020

Normal cellular and transformation-associated abl proteins share common sites for protein kinase C phosphorylation.

Viral transduction and chromosomal translocations of the c-abl gene result in the synthesis of abl proteins with structurally altered amino termini. These altered forms of the abl protein, but not the c-abl proteins, are detectably phosphorylated on tyrosine in vivo. In contrast, all forms of the abl protein are phosphorylated on serine following in vivo labeling with Pi. Treatment of NIH-3T3 cells with protein kinase C activators resulted in a four- to eightfold increase in the phosphorylation of murine c-abl due to modification of two serines on the c-abl protein. Purified protein kinase C phosphorylated all abl proteins at the same two sites. Both sites are precisely conserved in murine and human abl proteins. The sites on the abl proteins were found near the carboxy terminus. In contrast, for the epidermal growth factor receptor (T. Hunter, N. Ling, and J. A. Cooper, Nature [London] 311:480-483, 1984) and pp60src (K. L. Gould, J. R. Woodgett, J. A. Cooper, J. E. Buss, D. Shalloway, and T. Hunter, Cell 42:849-857, 1985), the sites of protein kinase C phosphorylation are amino-terminal to the kinase domain. The abl carboxy-terminal region is not necessary for the tyrosine kinase activity or transformation potential of the viral abl protein and may represent a regulatory domain. Using an in vitro immune complex kinase assay, we were not able to correlate reproducible changes in c-abl activity with phosphorylation by protein kinase C. However, the high degree of conservation of the phosphorylation sites for protein kinase C between human and mouse abl proteins suggests an important functional role.

Authors
Pendergast, AM; Traugh, JA; Witte, ON
MLA Citation
Pendergast, AM, Traugh, JA, and Witte, ON. "Normal cellular and transformation-associated abl proteins share common sites for protein kinase C phosphorylation." Mol Cell Biol 7.12 (December 1987): 4280-4289.
PMID
3125421
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
7
Issue
12
Publish Date
1987
Start Page
4280
End Page
4289

Regulation of phosphorylation of aminoacyl-tRNA synthetases in the high molecular weight core complex in reticulocytes.

Five aminoacyl-tRNA synthetases found in the high molecular weight core complex were phosphorylated in rabbit reticulocytes following labeling with 32P. The synthetases were isolated by affinity chromatography on tRNA-Sepharose followed by immunoprecipitation. The five synthetases phosphorylated were the glutamyl-, glutaminyl-, lysyl-, and aspartyl-tRNA synthetases and, to a lesser extent, the methionyl-tRNA synthetase. In addition, a 37,000-dalton protein, associated with the synthetase complex and tentatively identified as casein kinase I, was also phosphorylated in intact cells. Phosphoamino acid analysis of the proteins indicated all of the phosphate was on seryl residues. Incubation of reticulocytes with 32P in the presence of 8-bromo-cAMP and 3-isobutyl-1-methylxanthine resulted in a 6-fold increase in phosphorylation of the glutaminyl-tRNA synthetase and a 2-fold increase in phosphorylation of the aspartyl-tRNA synthetase. When the high molecular weight core complex was isolated by gel filtration/affinity chromatography, the profile of phosphorylation was similar to that observed by immunoprecipitation with a 9- and 3-fold stimulation of the glutaminyl- and aspartyl tRNA-synthetase, respectively. From this data it was concluded that the increased phosphorylation of the glutaminyl- and aspartyl-tRNA synthetases obtained with 8-bromo-cAMP did not appear to be involved in dissociation of the high molecular weight core complex.

Authors
Pendergast, AM; Venema, RC; Traugh, JA
MLA Citation
Pendergast, AM, Venema, RC, and Traugh, JA. "Regulation of phosphorylation of aminoacyl-tRNA synthetases in the high molecular weight core complex in reticulocytes." J Biol Chem 262.13 (May 5, 1987): 5939-5942.
PMID
2437110
Source
pubmed
Published In
The Journal of biological chemistry
Volume
262
Issue
13
Publish Date
1987
Start Page
5939
End Page
5942

Identification of three protein kinases which phosphorylate threonyl-tRNA synthetase from rat liver.

Threonyl-tRNA synthetase is phosphorylated in Chinese hamster ovary cells labeled with 32Pi [(1984) J. Biol. Chem. 259, 11160-11161]. Phosphorylation of the purified synthetase from rat liver has been examined with five different protein kinases. Three of the enzymes phosphorylate the synthetase, protease activated kinase I, the cAMP-dependent protein kinase, and the Ca2+, phospholipid-dependent protein kinase. Phosphorylation occurs exclusively on seryl residues. Two-dimensional phosphopeptide maps of tryptic digests of the phosphorylated synthetase are distinct with each protein kinase. These data suggest that multiple phosphorylation of the synthetase may occur in vivo.

Authors
Pendergast, AM; Traugh, JA
MLA Citation
Pendergast, AM, and Traugh, JA. "Identification of three protein kinases which phosphorylate threonyl-tRNA synthetase from rat liver." FEBS Lett 206.2 (October 6, 1986): 335-338.
PMID
3019777
Source
pubmed
Published In
FEBS Letters
Volume
206
Issue
2
Publish Date
1986
Start Page
335
End Page
338

Regulation of protein synthesis by phosphorylation of ribosomal protein S6 and aminoacyl-tRNA synthetases.

Authors
Traugh, JA; Pendergast, AM
MLA Citation
Traugh, JA, and Pendergast, AM. "Regulation of protein synthesis by phosphorylation of ribosomal protein S6 and aminoacyl-tRNA synthetases." Prog Nucleic Acid Res Mol Biol 33 (1986): 195-230. (Review)
PMID
3541042
Source
pubmed
Published In
Progress in nucleic acid research and molecular biology
Volume
33
Publish Date
1986
Start Page
195
End Page
230

Alteration of aminoacyl-tRNA synthetase activities by phosphorylation with casein kinase I.

The phosphorylation of a highly purified aminoacyl-tRNA synthetase complex from rabbit reticulocytes by the cyclic nucleotide-independent protein kinase, casein kinase I, has been examined, and the effects of phosphorylation on the synthetase activities were determined. The synthetase complex, purified as described (Kellermann, O., Tonetti, H., Brevet, A., Mirande, M., Pailliez, J.-P., and Waller, J.-P. (1982) J. Biol. Chem. 257, 11041-11048), contains seven aminoacyl-tRNA synthetases and four unidentified proteins and is free of endogenous protein kinase activity. Incubation of the complex with casein kinase I in the presence of ATP results in the phosphorylation of four synthetases, namely, glutamyl-, isoleucyl-, methionyl-, and lysyl-tRNA synthetases. Phosphorylation by casein kinase I alters binding of the aminoacyl-tRNA synthetase complex to tRNA-Sepharose. The phosphorylated synthetase complex elutes from tRNA-Sepharose at 190 mM NaCl, while the nonphosphorylated complex elutes at 275 mM NaCl. Phosphorylation by casein kinase I results in a significant inhibition of aminoacylation by the glutamyl-, isoleucyl-, methionyl-, and lysyl-tRNA synthetases; the activities of the nonphosphorylated synthetases remain unchanged. These data indicate that phosphorylation of aminoacyl-tRNA synthetases in the high molecular weight complex alters the activities of these enzymes. One of the unidentified proteins present in the complex (Mr 37,000) is also highly phosphorylated by casein kinase I. From a comparison of the properties and phosphopeptide pattern of this protein with that of casein kinase I, it appears that the Mr 37,000 protein in the synthetase complex is an inactive form of casein kinase I. This observation provides further evidence for a physiological role for casein kinase I in regulating synthetase activities.

Authors
Pendergast, AM; Traugh, JA
MLA Citation
Pendergast, AM, and Traugh, JA. "Alteration of aminoacyl-tRNA synthetase activities by phosphorylation with casein kinase I." J Biol Chem 260.21 (September 25, 1985): 11769-11774.
PMID
3862666
Source
pubmed
Published In
The Journal of biological chemistry
Volume
260
Issue
21
Publish Date
1985
Start Page
11769
End Page
11774

Phosphorylation of aminoacyl-tRNA synthetase complex from rabbit reticulocytes by casein kinase I

Authors
Pendergast, AM; Traugh, JA
MLA Citation
Pendergast, AM, and Traugh, JA. "Phosphorylation of aminoacyl-tRNA synthetase complex from rabbit reticulocytes by casein kinase I." Federation Proceedings 44.4 (1985): No.-3752.
Source
scival
Published In
Federation Proceedings
Volume
44
Issue
4
Publish Date
1985
Start Page
No.
End Page
3752

Laboratory practical on the preparation of mRNA-dependent rabbit reticulocyte lysate

Authors
Tomlinson, J; Pendergast, AM; Hronis, T; Jurnak, F
MLA Citation
Tomlinson, J, Pendergast, AM, Hronis, T, and Jurnak, F. "Laboratory practical on the preparation of mRNA-dependent rabbit reticulocyte lysate." Biochemical Education 12.1 (1984): 19-21.
Source
scival
Published In
Biochemical Education
Volume
12
Issue
1
Publish Date
1984
Start Page
19
End Page
21
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