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Boyce, Michael Scott

Positions:

Assistant Professor of Biochemistry

Biochemistry
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 2005

Ph.D. — Harvard Medical School

News:

1,100: Night Lights

October 16, 2013

Grants:

Cell signaling through O-GlcNAc reader proteins

Administered By
Biochemistry
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 2016
End Date
March 31, 2021

Organization and Function of Cellular Structure

Administered By
Basic Science Departments
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1975
End Date
June 30, 2020

Protein glycosylation in cancer cell signaling and metabolism

Administered By
Biochemistry
AwardedBy
Concern Foundation for Cancer Research
Role
Principal Investigator
Start Date
July 01, 2014
End Date
June 30, 2016

Publications:

The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor DELLA.

Plant development requires coordination among complex signaling networks to enhance the plant's adaptation to changing environments. DELLAs, transcription regulators originally identified as repressors of phytohormone gibberellin signaling, play a central role in integrating multiple signaling activities via direct protein interactions with key transcription factors. Here, we found that DELLA is mono-O-fucosylated by the novel O-fucosyltransferase SPINDLY (SPY) in Arabidopsis thaliana. O-fucosylation activates DELLA by promoting its interaction with key regulators in brassinosteroid- and light-signaling pathways, including BRASSINAZOLE-RESISTANT1 (BZR1), PHYTOCHROME-INTERACTING-FACTOR3 (PIF3) and PIF4. Moreover, spy mutants displayed elevated responses to gibberellin and brassinosteroid, and increased expression of common target genes of DELLAs, BZR1 and PIFs. Our study revealed that SPY-dependent protein O-fucosylation plays a key role in regulating plant development. This finding may have broader importance because SPY orthologs are conserved in prokaryotes and eukaryotes, thus suggesting that intracellular O-fucosylation may regulate a wide range of biological processes in diverse organisms.

Authors
Zentella, R; Sui, N; Barnhill, B; Hsieh, W-P; Hu, J; Shabanowitz, J; Boyce, M; Olszewski, NE; Zhou, P; Hunt, DF; Sun, T-P
MLA Citation
Zentella, R, Sui, N, Barnhill, B, Hsieh, W-P, Hu, J, Shabanowitz, J, Boyce, M, Olszewski, NE, Zhou, P, Hunt, DF, and Sun, T-P. "The Arabidopsis O-fucosyltransferase SPINDLY activates nuclear growth repressor DELLA." Nature chemical biology (February 28, 2017).
PMID
28244988
Source
epmc
Published In
Nature Chemical Biology
Publish Date
2017
DOI
10.1038/nchembio.2320

O -GlcNAcylation of master growth repressor DELLA by SECRET AGENT modulates multiple signaling pathways in Arabidopsis

Authors
Zentella, R; Hu, J; Hsieh, W-P; Matsumoto, PA; Dawdy, A; Barnhill, B; Oldenhof, H; Hartweck, LM; Maitra, S; Thomas, SG; Cockrell, S; Boyce, M; Shabanowitz, J; Hunt, DF; Olszewski, NE; Sun, T-P
MLA Citation
Zentella, R, Hu, J, Hsieh, W-P, Matsumoto, PA, Dawdy, A, Barnhill, B, Oldenhof, H, Hartweck, LM, Maitra, S, Thomas, SG, Cockrell, S, Boyce, M, Shabanowitz, J, Hunt, DF, Olszewski, NE, and Sun, T-P. "O -GlcNAcylation of master growth repressor DELLA by SECRET AGENT modulates multiple signaling pathways in Arabidopsis." Genes & Development 30.2 (January 15, 2016): 164-176.
Source
crossref
Published In
Genes & development
Volume
30
Issue
2
Publish Date
2016
Start Page
164
End Page
176
DOI
10.1101/gad.270587.115

A Chemical Glycoproteomics Platform Reveals O-GlcNAcylation of Mitochondrial Voltage-Dependent Anion Channel 2

Protein modification by O-linked β-N-acetylglucosamine (O-GlcNAc) is a critical cell signaling modality, but identifying signal-specific O-GlcNAcylation events remains a significant experimental challenge. Here, we describe a method for visualizing and analyzing organelle- and stimulus-specific O-GlcNAcylated proteins and use it to identify the mitochondrial voltage-dependent anion channel 2 (VDAC2) as an O-GlcNAc substrate. VDAC2-/- cells resist the mitochondrial dysfunction and apoptosis caused by global O-GlcNAc perturbation, demonstrating afunctional connection between O-GlcNAc signaling and mitochondrial physiology through VDAC2. More broadly, our method will enable the discovery of signal-specific O-GlcNAcylation events in a wide array of experimental contexts. © 2013 The Authors.

Authors
Palaniappan, KK; Hangauer, MJ; Smith, TJ; Smart, BP; Pitcher, AA; Cheng, EH; Bertozzi, CR; Boyce, M
MLA Citation
Palaniappan, KK, Hangauer, MJ, Smith, TJ, Smart, BP, Pitcher, AA, Cheng, EH, Bertozzi, CR, and Boyce, M. "A Chemical Glycoproteomics Platform Reveals O-GlcNAcylation of Mitochondrial Voltage-Dependent Anion Channel 2." Cell Reports 5.2 (2013): 546-552.
Source
scival
Published In
Cell Reports
Volume
5
Issue
2
Publish Date
2013
Start Page
546
End Page
552
DOI
10.1016/j.celrep.2013.08.048

Erratum: Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury (Nature Chemical Biology (2005) 1 (112-119))

Authors
Degterev, A; Huang, Z; Boyce, M; Li, Y; Jagtap, P; Mizushima, N; Cuny, GD; Mitchison, TJ; Moskowitz, MA; Yuan, J
MLA Citation
Degterev, A, Huang, Z, Boyce, M, Li, Y, Jagtap, P, Mizushima, N, Cuny, GD, Mitchison, TJ, Moskowitz, MA, and Yuan, J. "Erratum: Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury (Nature Chemical Biology (2005) 1 (112-119))." Nature Chemical Biology 9.3 (2013): 192--.
Source
scival
Published In
Nature Chemical Biology
Volume
9
Issue
3
Publish Date
2013
Start Page
192-
DOI
10.1038/nchembio0313-192a

A Chemical Glycoproteomics Platform Reveals O-GlcNAcylation of Mitochondrial Voltage-Dependent Anion Channel 2

Protein modification by O-linked β-N-acetylglucosamine (O-GlcNAc) is a critical cell signaling modality, but identifying signal-specific O-GlcNAcylation events remains a significant experimental challenge. Here, we describe a method for visualizing and analyzing organelle- and stimulus-specific O-GlcNAcylated proteins and use it to identify the mitochondrial voltage-dependent anion channel 2 (VDAC2) as an O-GlcNAc substrate. VDAC2 -/- cells resist the mitochondrial dysfunction and apoptosis caused by global O-GlcNAc perturbation, demonstrating a functional connection between O-GlcNAc signaling and mitochondrial physiology through VDAC2. More broadly, our method will enable the discovery of signal-specific O-GlcNAcylation events in a wide array of experimental contexts. © 2013 The Authors.

Authors
Palaniappan, KK; Hangauer, MJ; Smith, TJ; Smart, BP; Pitcher, AA; Cheng, EH; Bertozzi, CR; Boyce, M
MLA Citation
Palaniappan, KK, Hangauer, MJ, Smith, TJ, Smart, BP, Pitcher, AA, Cheng, EH, Bertozzi, CR, and Boyce, M. "A Chemical Glycoproteomics Platform Reveals O-GlcNAcylation of Mitochondrial Voltage-Dependent Anion Channel 2." Cell Reports (2013).
PMID
24120863
Source
scival
Published In
Cell Reports
Publish Date
2013
DOI
10.1016/j.celrep.2013.08.048

Metabolic labeling enables selective photocrosslinking of O-GlcNAc-modified proteins to their binding partners

O-linked β-N-acetylglucosamine (O-GlcNAc) is a reversible posttranslational modification found on hundreds of nuclear and cytoplasmic proteins in higher eukaryotes. Despite its ubiquity and essentiality in mammals, functional roles for the O-GlcNAc modification remain poorly defined. Here we develop a combined genetic and chemical approach that enables introduction of the diazirine photocrosslinker onto the O-GlcNAc modification in cells. We engineered mammalian cells to produce diazirine-modified O-GlcNAc by expressing a mutant form of UDP-GlcNAc pyrophosphorylase and subsequently culturing these cells with a cell-permeable, diazirine-modified form of GlcNAc-1-phosphate. Irradiation of cells with UV light activated the crosslinker, resulting in formation of covalent bonds between O-GlcNAc-modified proteins and neighboring molecules, which could be identified by mass spectrometry. We used this method to identify interaction partners for the O-GlcNAc-modified FG-repeat nucleoporins. We observed crosslinking between FG-repeat nucleoporins and nuclear transport factors, suggesting that O-GlcNAc residues are intimately associated with essential recognition events in nuclear transport. Further, we propose that the method reported here could find widespread use in investigating the functional consequences of O-GlcNAcylation.

Authors
Yu, S-H; Boyce, M; Wands, AM; Bond, MR; Bertozzi, CR; Kohler, JJ
MLA Citation
Yu, S-H, Boyce, M, Wands, AM, Bond, MR, Bertozzi, CR, and Kohler, JJ. "Metabolic labeling enables selective photocrosslinking of O-GlcNAc-modified proteins to their binding partners." Proceedings of the National Academy of Sciences of the United States of America 109.13 (2012): 4834-4839.
PMID
22411826
Source
scival
Published In
Proceedings of the National Academy of Sciences of USA
Volume
109
Issue
13
Publish Date
2012
Start Page
4834
End Page
4839
DOI
10.1073/pnas.1114356109

Endoplasmic reticulum stress response in cell death and cell survival

© Cambridge University Press 2011.Introduction The endoplasmic reticulum (ER) serves as the primary cellular protein processing factory where polypeptides destined for secretion or membrane insertion are folded. This membrane-bound organelle recruits translating ribosomes, translocates newly synthesized peptides into its lumen, and promotes a variety of post-translational modifications and chaperone-facilitated folding events. Additionally, in higher eukaryotes ER serves as the major intracellular Ca2+store. Because the ER encompasses about half the total membrane area and one-third the newly translated proteins in a typical eukaryotic cell, its proper function is critical for numerous aspects of cell physiology, including vesicle trafficking, lipid and membrane biogenesis, and protein targeting and secretion. Accordingly, metazoan cells react rapidly to ER dysfunction through a set of adaptive pathways known collectively as the ER stress response (ESR).

Authors
Boyce, M; Lipinski, MM; Py, BF; Yuan, J
MLA Citation
Boyce, M, Lipinski, MM, Py, BF, and Yuan, J. "Endoplasmic reticulum stress response in cell death and cell survival." (January 1, 2011): 51-62. (Chapter)
Source
scopus
Publish Date
2011
Start Page
51
End Page
62
DOI
10.1017/CBO9780511976094.006

Bringing chemistry to life

Authors
Boyce, M; Bertozzi, CR
MLA Citation
Boyce, M, and Bertozzi, CR. "Bringing chemistry to life." Nature Methods 8.8 (2011): 638-642.
PMID
21799498
Source
scival
Published In
Nature Methods
Volume
8
Issue
8
Publish Date
2011
Start Page
638
End Page
642
DOI
10.1038/nmeth.1657

Cell surface glycoproteomic analysis of prostate cancer-derived PC-3 cells

Most clinically approved biomarkers of cancer are glycoproteins, and those residing on the cell surface are of particular interest in biotherapeutics. We report a method for selective labeling, affinity enrichment, and identification of cell-surface glycoproteins. PC-3 cells and primary human prostate cancer tissue were treated with peracetylated N-azidoacetylgalactosamine, resulting in metabolic labeling of cell surface glycans with the azidosugar. We used mass spectrometry to identify over 70 cell surface glycoproteins and biochemically validated CD146 and integrin beta-4, both of which are known to promote metastatic behavior. These results establish cell-surface glycoproteomics as an effective technique for discovery of cancer biomarkers. © 2011 Elsevier Ltd. All rights reserved.

Authors
Hubbard, SC; Boyce, M; McVaugh, CT; Peehl, DM; Bertozzi, CR
MLA Citation
Hubbard, SC, Boyce, M, McVaugh, CT, Peehl, DM, and Bertozzi, CR. "Cell surface glycoproteomic analysis of prostate cancer-derived PC-3 cells." Bioorganic and Medicinal Chemistry Letters 21.17 (2011): 4945-4950.
PMID
21798741
Source
scival
Published In
Bioorganic & Medicinal Chemistry Letters
Volume
21
Issue
17
Publish Date
2011
Start Page
4945
End Page
4950
DOI
10.1016/j.bmcl.2011.05.045

Metabolic cross-talk allows labeling of O-linked β-N- acetylglucosamine-modified proteins via the N-acetylgalactosamine salvage pathway

Hundreds of mammalian nuclear and cytoplasmic proteins are reversibly glycosylated by O-linked β-N-acetylglucosamine (O-GlcNAc) to regulate their function, localization, and stability. Despite its broad functional significance, the dynamic and posttranslational nature of O-GlcNAc signaling makes it challenging to study using traditional molecular and cell biological techniques alone. Here, we report that metabolic cross-talk between the N-acetylgalactosamine salvage and O-GlcNAcylation pathways can be exploited for the tagging and identification of O-GlcNAcylated proteins. We found that N-azidoacetylgalactosamine (GalNAz) is converted by endogenous mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to UDP-N- azidoacetylglucosamine (GlcNAz). O-GlcNAc transferase accepts UDP-GlcNAz as a nucleotide-sugar donor, appending an azidosugar onto its native substrates, which can then be detected by covalent labeling using azide-reactive chemical probes. In a proof-of-principle proteomics experiment, we used metabolic GalNAz labeling of human cells and a bioorthogonal chemical probe to affinity-purify and identify numerous O-GlcNAcylated proteins. Our work provides a blueprint for a wide variety of future chemical approaches to identify, visualize, and characterize dynamic O-GlcNAc signaling.

Authors
Boyce, M; Carrico, IS; Ganguli, AS; Yu, S-H; Hangauer, MJ; Hubbard, SC; Kohler, JJ; Bertozzi, CR
MLA Citation
Boyce, M, Carrico, IS, Ganguli, AS, Yu, S-H, Hangauer, MJ, Hubbard, SC, Kohler, JJ, and Bertozzi, CR. "Metabolic cross-talk allows labeling of O-linked β-N- acetylglucosamine-modified proteins via the N-acetylgalactosamine salvage pathway." Proceedings of the National Academy of Sciences of the United States of America 108.8 (2011): 3141-3146.
PMID
21300897
Source
scival
Published In
Proceedings of the National Academy of Sciences of USA
Volume
108
Issue
8
Publish Date
2011
Start Page
3141
End Page
3146
DOI
10.1073/pnas.1010045108

A Chemical Method for Labeling Lysine Methyltransferase Substrates

Several protein lysine methyltransferases (PKMTs) modify histones to regulate chromatin-dependent cellular processes, such as transcription, DNA replication and DNA damage repair. PKMTs are likely to have many additional substrates in addition to histones, but relatively few nonhistone substrates have been characterized, and the substrate specificity for many PKMTs has yet to be defined. Thus, new unbiased methods are needed to find PKMT substrates. Here, we describe a chemical biology approach for unbiased, proteome-wide identification of novel PKMT substrates. Our strategy makes use of an alkyne-bearing S-adenosylmethionine (SAM) analogue, which is accepted by the PKMT, SETDB1, as a cofactor, resulting in the enzymatic attachment of a terminal alkyne to its substrate. Such labeled proteins can then be treated with azide-functionalized probes to ligate affinity handles or fluorophores to the PKMT substrates. As a proof-of-concept, we have used SETDB1 to transfer the alkyne moiety from the SAM analogue onto a recombinant histone H3 substrate. We anticipate that this chemical method will find broad use in epigenetics to enable unbiased searches for new PKMT substrates by using recombinant enzymes and unnatural SAM cofactors to label and purify many substrates simultaneously from complex organelle or cell extracts.Click art: The transfer of an alkyne moiety from an S-adenosylmethionine analogue (1) onto a lysine residue by the methyltransferase, KMT, allowed the tagging of the modified substrate for subsequent analysis by copper-catalyzed click chemistry, as demonstrated in the scheme. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Authors
Binda, O; Boyce, M; Rush, JS; Palaniappan, KK; Bertozzi, CR; Gozani, O
MLA Citation
Binda, O, Boyce, M, Rush, JS, Palaniappan, KK, Bertozzi, CR, and Gozani, O. "A Chemical Method for Labeling Lysine Methyltransferase Substrates." ChemBioChem 12.2 (2011): 330-334.
PMID
21243721
Source
scival
Published In
Chembiochem
Volume
12
Issue
2
Publish Date
2011
Start Page
330
End Page
334
DOI
10.1002/cbic.201000433

A critical role of eEF-2K in mediating autophagy in response to multiple cellular stresses

The phosphorylation of the subunit α of eukaryotic translation initiation factor 2 (eIF2α), a critical regulatory event in controlling protein translation, has recently been found to mediate the induction of autophagy. However, the mediators of autophagy downstream of eIF2α remain unknown. Here, we provide evidence that eIF2α phosphorylation is required for phosphorylation of eukaryotic elongation factor 2 (eEF-2) during nutrient starvation. In addition, we show that eukaryotic elongation factor 2 kinase (eEF-2K) is also required for autophagy signaling during ER stress, suggesting that phosphorylation of eEF-2 may serve as an integrator of various cell stresses for autophagy signaling. On the other hand, although the activation of eEF-2K in response to starvation requires the phosphorylation of eIF2α, additional pathways relying partly on Ca2+ flux may control eEF-2K activity during ER stress, as eIF2α phosphorylation is dispensable for both eEF-2 phosphorylation and autophagy in this context. © 2009 Landes Bioscience.

Authors
Bénédicte, F; Boyce, M; Yuan, J
MLA Citation
Bénédicte, F, Boyce, M, and Yuan, J. "A critical role of eEF-2K in mediating autophagy in response to multiple cellular stresses." Autophagy 5.3 (2009): 393-396.
PMID
19221463
Source
scival
Published In
Autophagy
Volume
5
Issue
3
Publish Date
2009
Start Page
393
End Page
396
DOI
10.4161/auto.5.3.7762

ICP34.5-dependent and -independent activities of salubrinal in herpes simplex virus-1 infected cells

The small molecule salubrinal has antiviral activity against herpes simplex virus-1 (HSV-1) and inhibits dephosphorylation of eIF2α mediated by the HSV-1 protein ICP34.5. We investigated whether salubrinal's activities in infected cells depend on ICP34.5. An ICP34.5 deletion mutant was as sensitive as wild type HSV-1 to salubrinal inhibition of plaque formation in Vero cells. However, salubrinal induced formation of syncytia in infected Vero cells, which was enhanced by ICP34.5 mutations. Expression of HSV-1 US11 with immediate early kinetics, which is known to suppress the effects of ICP34.5 mutations, resulted in slight resistance to salubrinal in murine embryonic fibroblasts, and substantial resistance in those cells when ICP34.5 was additionally mutated. ICP34.5 mutations, but not immediate early expression of US11, prevented salubrinal's ability to increase phosphorylation of eIF2α during HSV-1 infection of Vero cells. Taken together, our data indicate that salubrinal has both ICP34.5-dependent and -independent activities in HSV-1 infected cells. © 2008 Elsevier Inc. All rights reserved.

Authors
Bryant, KF; Macari, ER; Malik, N; Boyce, M; Yuan, J; Coen, DM
MLA Citation
Bryant, KF, Macari, ER, Malik, N, Boyce, M, Yuan, J, and Coen, DM. "ICP34.5-dependent and -independent activities of salubrinal in herpes simplex virus-1 infected cells." Virology 379.2 (2008): 197-204.
PMID
18684481
Source
scival
Published In
Virology
Volume
379
Issue
2
Publish Date
2008
Start Page
197
End Page
204
DOI
10.1016/j.virol.2008.06.028

A pharmacoproteomic approach implicates eukaryotic elongation factor 2 kinase in ER stress-induced cell death

Apoptosis triggered by endoplasmic reticulum (ER) stress has been implicated in many diseases but its cellular regulation remains poorly understood. Previously, we identified salubrinal (sal), a small molecule that protects cells from ER stress-induced apoptosis by selectively activating a subset of endogenous ER stress-signaling events. Here, we use sal as a probe in a proteomic approach to discover new information about the endogenous cellular response to ER stress. We show that sal induces phosphorylation of the translation elongation factor eukaryotic translation elongation factor 2 (eEF-2), an event that depends on eEF-2 kinase (eEF-2K). ER stress itself also induces eEF-2K-dependent eEF-2 phosphorylation, and this pathway promotes translational arrest and cell death in this context, identifying eEF-2K as a hitherto unknown regulator of ER stress-induced apoptosis. Finally, we use both sal and ER stress models to show that eEF-2 phosphorylation can be activated by at least two signaling mechanisms. Our work identifies eEF-2K as a new component of the ER stress response and underlines the utility of novel small molecules in discovering new cell biology.

Authors
Boyce, M; Py, BF; Ryazanov, AG; Minden, JS; Long, K; Ma, D; Yuan, J
MLA Citation
Boyce, M, Py, BF, Ryazanov, AG, Minden, JS, Long, K, Ma, D, and Yuan, J. "A pharmacoproteomic approach implicates eukaryotic elongation factor 2 kinase in ER stress-induced cell death." Cell Death and Differentiation 15.3 (2008): 589-599.
PMID
18188169
Source
scival
Published In
Cell Death & Differentiation
Volume
15
Issue
3
Publish Date
2008
Start Page
589
End Page
599
DOI
10.1038/sj.cdd.4402296

A genome-wide RNAi screen reveals multiple regulators of caspase activation

Apoptosis is an evolutionally conserved cellular suicide mechanism that can be activated in response to a variety of stressful stimuli. Increasing evidence suggests that apoptotic regulation relies on specialized cell death signaling pathways and also integrates diverse signals from additional regulatory circuits, including those of cellular homeostasis. We present a genome-wide RNA interference screen to systematically identify regulators of apoptosis induced by DNA damage in Drosophila melanogaster cells. We identify 47 double-stranded RNA that target a functionally diverse set of genes, including several with a known function in promoting cell death. Further characterization uncovers 10 genes that influence caspase activation upon the removal of Drosophila inhibitor of apoptosis 1. This set includes the Drosophila initiator caspase Dronc and, surprisingly, several metabolic regulators, a candidate tumor suppressor, Charlatan, and an N-acetyltransferase, ARD1. Importantly, several of these genes show functional conservation in regulating apoptosis in mammalian cells. Our data suggest a previously unappreciated fundamental connection between various cellular processes and caspase-dependent cell death. © The Rockefeller University Press.

Authors
Yi, CH; Sogah, DK; Boyce, M; Degterev, A; Christofferson, DE; Yuan, J
MLA Citation
Yi, CH, Sogah, DK, Boyce, M, Degterev, A, Christofferson, DE, and Yuan, J. "A genome-wide RNAi screen reveals multiple regulators of caspase activation." Journal of Cell Biology 179.4 (2007): 619-626.
PMID
17998402
Source
scival
Published In
The Journal of Cell Biology
Volume
179
Issue
4
Publish Date
2007
Start Page
619
End Page
626
DOI
10.1083/jcb.200708090

Selective inhibition of eukaryotic translation initiation factor 2α dephosphorylation potentiates fatty acid-induced endoplasmic reticulum stress and causes pancreatic β-cell dysfunction and apoptosis

Free fatty acids cause pancreatic β-cell apoptosis and may contribute to β-cell loss in type 2 diabetes via the induction of endoplasmic reticulum stress. Reductions in eukaryotic translation initiation factor (eIF) 2α phosphorylation trigger β-cell failure and diabetes. Salubrinal selectively inhibits eIF2α dephosphorylation, protects other cells against endoplasmic reticulum stress-mediated apoptosis, and has been proposed as a β-cell protector. Unexpectedly, salubrinal induced apoptosis in primary β-cells, and it potentiated the deleterious effects of oleate and palmitate. Salubrinal induced a marked eIF2α phosphorylation and potentiated the inhibitory effects of free fatty acids on protein synthesis and insulin release. The synergistic activation of the PERK-eIF2α branch of the endoplasmic reticulum stress response, but not of the IRE1 and activating transcription factor-6 pathways, led to a marked induction of activating transcription factor-4 and the pro-apoptotic transcription factor CHOP. Our findings demonstrate that excessive eIF2α phosphorylation is poorly tolerated by β-cells and exacerbates free fatty acid-induced apoptosis. This modifies the present paradigm regarding the beneficial role of eIF2α phosphorylation in β-cells and must be taken into consideration when designing therapies to protect β-cells in type 2 diabetes. © 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Authors
Cnop, M; Ladriere, L; Hekerman, P; Ortis, F; Cardozo, AK; Dogusan, Z; Flamez, D; Boyce, M; Yuan, J; Eizirik, DL
MLA Citation
Cnop, M, Ladriere, L, Hekerman, P, Ortis, F, Cardozo, AK, Dogusan, Z, Flamez, D, Boyce, M, Yuan, J, and Eizirik, DL. "Selective inhibition of eukaryotic translation initiation factor 2α dephosphorylation potentiates fatty acid-induced endoplasmic reticulum stress and causes pancreatic β-cell dysfunction and apoptosis." Journal of Biological Chemistry 282.6 (2007): 3989-3997.
PMID
17158450
Source
scival
Published In
The Journal of biological chemistry
Volume
282
Issue
6
Publish Date
2007
Start Page
3989
End Page
3997
DOI
10.1074/jbc.M607627200

Cellular response to endoplasmic reticulum stress: A matter of life or death

The proper functioning of the endoplasmic reticulum (ER) is critical for numerous aspects of cell physiology. Accordingly, all eukaryotes react rapidly to ER dysfunction through a set of adaptive pathways known collectively as the ER stress response (ESR). Normally, this suite of responses succeeds in restoring ER homeostasis. However, in metazoans, persistent or intense ER stress can also trigger programmed cell death, or apoptosis. ER stress and the apoptotic program coupled to it have been implicated in many important pathologies but the regulation and execution of ER stress-induced apoptosis in mammals remain incompletely understood. Here, we review what is known about the ESR in both yeast and mammals, and highlight recent findings on the mechanism and pathophysiological importance of ER stress-induced apoptosis. © 2006 Nature Publishing Group. All rights reserved.

Authors
Boyce, M; Yuan, J
MLA Citation
Boyce, M, and Yuan, J. "Cellular response to endoplasmic reticulum stress: A matter of life or death." Cell Death and Differentiation 13.3 (2006): 363-373.
PMID
16397583
Source
scival
Published In
Cell Death & Differentiation
Volume
13
Issue
3
Publish Date
2006
Start Page
363
End Page
373
DOI
10.1038/sj.cdd.4401817

Base-pairing potential identified by in vitro selection predicts the kinked RNA backbone observed in the crystal structure of the Alfalfa Mosaic virus RNA-coat protein complex

The three-dimensional structure of the 3′ terminus of alfalfa mosaic virus RNA in complex with an amino-terminal coat protein peptide revealed an unusual RNA fold with inter-AUGC basepairing stabilized by key arginine residues (Guogas, et al., 2004). To probe viral RNA interactions with the full-length coat protein, we have used in vitro genetic selection to characterize potential folding patterns among RNAs isolated from a complex randomized pool. Nitrocellulose filter retention, electrophoretic mobility bandshift analysis, and hydroxyl radical footprinting techniques were used to define binding affinities and to localize the potential RNA-protein interaction sites. Minimized binding sites were identified that included both the randomized domain and a portion of the constant regions of the selected RNAs. The selected RNAs, identified by their ability to bind full-length coat protein, have the potential to form the same unusual inter-AUGC Watson-Crick base pairs observed in the crystal structure, although the primary sequences diverge from the wild-type RNA. A constant feature of both the wild-type RNA and the selected RNAs is a G ribonucleotide in the third position of an AUGC-like repeat. Competitive binding assays showed that substituting adenosine for the constant guanosine in either the wild-type or selected RNAs impaired coat protein binding. These data suggest that the interactions observed in the RNA-peptide structure are likely recapitulated when the full-length protein binds. Further, the results underscore the power of in vitro genetic selection for probing RNA-protein structure and function. Copyright © 2005 John Wiley & Sons, Ltd.

Authors
Boyce, M; Scott, F; Guogas, LM; Gehrke, L
MLA Citation
Boyce, M, Scott, F, Guogas, LM, and Gehrke, L. "Base-pairing potential identified by in vitro selection predicts the kinked RNA backbone observed in the crystal structure of the Alfalfa Mosaic virus RNA-coat protein complex." Journal of Molecular Recognition 19.1 (2006): 68-78.
PMID
16312015
Source
scival
Published In
Journal of Molecular Recognition
Volume
19
Issue
1
Publish Date
2006
Start Page
68
End Page
78
DOI
10.1002/jmr.759

Corrigendum: Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury

Authors
Degterev, A; Huang, Z; Boyce, M; Li, Y; Jagtap, P; Mizushima, N; Cuny, GD; Mitchison, TJ; Moskowitz, MA; Yuan, J
MLA Citation
Degterev, A, Huang, Z, Boyce, M, Li, Y, Jagtap, P, Mizushima, N, Cuny, GD, Mitchison, TJ, Moskowitz, MA, and Yuan, J. "Corrigendum: Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury." Nature Chemical Biology 1.4 (September 2005): 234-234.
Source
crossref
Published In
Nature Chemical Biology
Volume
1
Issue
4
Publish Date
2005
Start Page
234
End Page
234
DOI
10.1038/nchembio0905-234a

Structure-activity relationship studies of salubrinal lead to its active biotinylated derivative

The synthesis and structure-activity relationships (SAR) of salubrinal, a small molecule that protects cells from apoptosis induced by endoplasmic reticulum (ER) stress, are described. It is revealed that the trichloromethyl group greatly contributes to the activity. Based on the SAR results, salubrinal was converted into a biotinylated derivative which retains activity and can be used as a biological tool for target identification. © 2005 Elsevier Ltd. All rights reserved.

Authors
Long, K; Boyce, M; Lin, H; Yuan, J; Ma, D
MLA Citation
Long, K, Boyce, M, Lin, H, Yuan, J, and Ma, D. "Structure-activity relationship studies of salubrinal lead to its active biotinylated derivative." Bioorganic and Medicinal Chemistry Letters 15.17 (2005): 3849-3852.
PMID
16002288
Source
scival
Published In
Bioorganic and Medicinal Chemistry Letters
Volume
15
Issue
17
Publish Date
2005
Start Page
3849
End Page
3852
DOI
10.1016/j.bmcl.2005.05.120

Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury.

The mechanism of apoptosis has been extensively characterized over the past decade, but little is known about alternative forms of regulated cell death. Although stimulation of the Fas/TNFR receptor family triggers a canonical 'extrinsic' apoptosis pathway, we demonstrated that in the absence of intracellular apoptotic signaling it is capable of activating a common nonapoptotic death pathway, which we term necroptosis. We showed that necroptosis is characterized by necrotic cell death morphology and activation of autophagy. We identified a specific and potent small-molecule inhibitor of necroptosis, necrostatin-1, which blocks a critical step in necroptosis. We demonstrated that necroptosis contributes to delayed mouse ischemic brain injury in vivo through a mechanism distinct from that of apoptosis and offers a new therapeutic target for stroke with an extended window for neuroprotection. Our study identifies a previously undescribed basic cell-death pathway with potentially broad relevance to human pathologies.

Authors
Degterev, A; Huang, Z; Boyce, M; Li, Y; Jagtap, P; Mizushima, N; Cuny, GD; Mitchison, TJ; Moskowitz, MA; Yuan, J
MLA Citation
Degterev, A, Huang, Z, Boyce, M, Li, Y, Jagtap, P, Mizushima, N, Cuny, GD, Mitchison, TJ, Moskowitz, MA, and Yuan, J. "Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury." Nature chemical biology 1.2 (2005): 112-119.
PMID
16408008
Source
scival
Published In
Nature Chemical Biology
Volume
1
Issue
2
Publish Date
2005
Start Page
112
End Page
119
DOI
10.1038/nchembio711

A selective inhibitor of elF2α dephosphorylation protects cells from ER stress

Most protein phosphatases have little intrinsic substrate specificity, making selective pharmacological inhibition of specific dephosphorylation reactions a challenging problem. In a screen for small molecules that protect cells from endoplasmic reticulum (ER) stress, we identified salubrinal, a selective inhibitor of cellular complexes that dephosphorylate eukaryotic translation initiation factor 2 subunit α (elF2α). Salubrinal also blocks elF2α dephosphorylation mediated by a herpes simplex virus protein and inhibits viral replication. These results suggest that selective chemical inhibitors of elF2α dephosphorylation may be useful in diseases involving ER stress or viral infection. More broadly, salubrinal demonstrates the feasibility of selective pharmacological targeting of cellular dephosphorylation events.

Authors
Boyce, M; Bryant, KF; Jousse, C; Long, K; Harding, HP; Scheuner, D; Kaufman, RJ; Ma, D; Coen, DM; Ron, D; Yuan, J
MLA Citation
Boyce, M, Bryant, KF, Jousse, C, Long, K, Harding, HP, Scheuner, D, Kaufman, RJ, Ma, D, Coen, DM, Ron, D, and Yuan, J. "A selective inhibitor of elF2α dephosphorylation protects cells from ER stress." Science 307.5711 (2005): 935-939.
PMID
15705855
Source
scival
Published In
Science
Volume
307
Issue
5711
Publish Date
2005
Start Page
935
End Page
939
DOI
10.1126/science.1101902

Caspases: An ancient cellular sword of Damocles

Caspases are a family of cysteine proteases homologous to the Caenorhabditis elegans programmed cell death gene product CED-3. Caspases and their distant relatives, meta- and paracaspases, have been found in phylogenetically distant nonmetazoan groups, including plants, fungi and prokaryotes. This review summarizes the current information on the mechanisms and functions of non-mammalian caspases and their relatives in apoptotic and nonapoptotic processes, and explores the possible evolutionary origin of the caspase family.

Authors
Boyce, M; Degterev, A; Yuan, J
MLA Citation
Boyce, M, Degterev, A, and Yuan, J. "Caspases: An ancient cellular sword of Damocles." Cell Death and Differentiation 11.1 (2004): 29-37.
PMID
14647235
Source
scival
Published In
Cell Death and Differentiation
Volume
11
Issue
1
Publish Date
2004
Start Page
29
End Page
37
DOI
10.1038/sj.cdd.4401339

A decade of caspases

Caspases are a family of cysteine proteases that play important roles in regulating apoptosis. A decade of research has generated a wealth of information on the signal transduction pathways mediated by caspases, the distinct functions of individual caspases and the mechanisms by which caspases mediate apoptosis and a variety of physiological and pathological processes.

Authors
Degterev, A; Boyce, M; Yuan, J
MLA Citation
Degterev, A, Boyce, M, and Yuan, J. "A decade of caspases." Oncogene 22.53 REV. ISS. 7 (2003): 8543-8567.
PMID
14634618
Source
scival
Published In
Oncogene
Volume
22
Issue
53 REV. ISS. 7
Publish Date
2003
Start Page
8543
End Page
8567
DOI
10.1038/sj.onc.1207107

Life and death in paradise

Over 500 researchers participated in a recent American Association for Cancer Research special conference, entitled "Apoptosis and Cancer: Basic Mechanisms and Therapeutic Opportunities in the Post-Genomic Era" (February 13-17, 2002) in sunny Hawaii (Hilton Waikoloa village, Kona, Hawaii). The meeting participants presented the most recent findings on the mechanisms regulating cell death in cancer. In the past decade, apoptosis research has undergone a quantum leap, metamorphosing from a descriptive, phenomenological discipline into a molecularly defined, highly complex signalling field. This transformation was highlighted in the conference's opening talk by meeting co-organizer, John Reed (The Burnham Institute, La Jolla, CA). Reed and colleagues used published protein functional information and bio-informatic mining of the available human genome databases to tabulate the number of human proteins predicted to be involved in regulating apoptosis. The list includes 11 catalytically active caspases, 26 CARD (caspase associated recruitment domain)-, 32 DD (death domain)-, 12 DED (death effector domain)-, 8 BIR (baculovirus inhibitor of apoptosis protein region)-, 24 BH (Bcl-2 homology)-, and 34 PAAD/PYD (pyrin/PAAD)-containing sequences.

Authors
Gozani, O; Boyce, M; Yoo, L; Karuman, P; Yuan, J
MLA Citation
Gozani, O, Boyce, M, Yoo, L, Karuman, P, and Yuan, J. "Life and death in paradise." Nature Cell Biology 4.6 (2002): E159-E162.
PMID
12042835
Source
scival
Published In
Nature Cell Biology
Volume
4
Issue
6
Publish Date
2002
Start Page
E159
End Page
E162
DOI
10.1038/ncb0602-e159

The channel of death

The proapoptotic members of the Bcl-2 family have been proposed to participate in the formation of a channel that releases these apoptogenic factors when mitochondria receive apoptotic signals. A recent study provides the first direct, biophysical measurement of a potentially apoptosis-specific mitochondrial channel, which is regulated by Bcl-2 family members and may play a primary role in the release of the proapoptotic factors.

Authors
Degterev, A; Boyce, M; Yuan, J
MLA Citation
Degterev, A, Boyce, M, and Yuan, J. "The channel of death." Journal of Cell Biology 155.5 (2001): 695-697.
PMID
11724809
Source
scival
Published In
Journal of Cell Biology
Volume
155
Issue
5
Publish Date
2001
Start Page
695
End Page
697
DOI
10.1083/jcb.200110147
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