A strategy for modulation of enzymes in the ubiquitin system.
Andreas Ernst,George V. Avvakumov,Jiefei Tong,Yihui Fan,Yanling Zhao,Philipp Alberts,Avinash Persaud,John R. Walker,Ana-Mirela Neculai,Dante Neculai,Andrew Vorobyov,Pankaj Garg,Linda G. Beatty,Pak-Kei Chan,Yu Chi Juang,Marie-Claude Landry,Christina Yeh,Christina Yeh,Elton Zeqiraj,Konstantina Karamboulas,Abdellah Allali-Hassani,Masoud Vedadi,Mike Tyers,Mike Tyers,Jason Moffat,Frank Sicheri,Frank Sicheri,Laurence Pelletier,Laurence Pelletier,Daniel Durocher,Daniel Durocher,Brian Raught,Daniela Rotin,Jianhua Yang,Michael Moran,Sirano Dhe-Paganon,Sirano Dhe-Paganon,Sachdev S. Sidhu +37 more
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TLDR
This work used massively diverse combinatorial libraries of ubiquitin variants to develop inhibitors of four deubiquitinases (DUBs) and analyzed the DUB-inhibitor complexes with crystallography to report a method to target the myriad enzymes that govern ubiquitination of protein substrates.Abstract:
The ubiquitin system regulates virtually all aspects of cellular function. We report a method to target the myriad enzymes that govern ubiquitination of protein substrates. We used massively diverse combinatorial libraries of ubiquitin variants to develop inhibitors of four deubiquitinases (DUBs) and analyzed the DUB-inhibitor complexes with crystallography. We extended the selection strategy to the ubiquitin conjugating (E2) and ubiquitin ligase (E3) enzymes and found that ubiquitin variants can also enhance enzyme activity. Last, we showed that ubiquitin variants can bind selectively to ubiquitin-binding domains. Ubiquitin variants exhibit selective function in cells and thus enable orthogonal modulation of specific enzymatic steps in the ubiquitin system.read more
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Emerging Potential of Therapeutic Targeting of Ubiquitin-Specific Proteases in the Treatment of Cancer
TL;DR: The ubiquitin-proteasome system (UPS) has emerged as a therapeutic focus and target for the treatment of cancer as discussed by the authors, and targeting deubiquitinases (DUBs) is a promising approach in regulation of the UPS.
Journal ArticleDOI
System-Wide Modulation of HECT E3 Ligases with Selective Ubiquitin Variant Probes
Wei Zhang,Kuen-Phon Wu,Maria A. Sartori,Hari B. Kamadurai,Alban Ordureau,Chong Jiang,Peter Y. Mercredi,Ryan Murchie,Jicheng Hu,Avinash Persaud,Manjeet Mukherjee,Nan Li,Anne Doye,John R. Walker,Yi Sheng,Zhenyue Hao,Yanjun Li,Kevin R. Brown,Emmanuel Lemichez,Junjie Chen,Yufeng Tong,Yufeng Tong,J. Wade Harper,Jason Moffat,Daniela Rotin,Brenda A. Schulman,Sachdev S. Sidhu +26 more
TL;DR: This work demonstrates versatility of UbVs for modulating activity across an E3 family, defines mechanisms and provides a toolkit for probing functions of HECT E3s, and establishes a general strategy for systematic development of modulators targeting families of signaling proteins.
Journal ArticleDOI
Systematic approaches to identify E3 ligase substrates.
TL;DR: Systematic approaches to identify and validate UPS targets are highlighted and how they are underpinning rapid advances in the understanding of the biochemistry and biology of the UPS are discussed.
Journal ArticleDOI
Dynamics of PARKIN-Dependent Mitochondrial Ubiquitylation in Induced Neurons and Model Systems Revealed by Digital Snapshot Proteomics
Alban Ordureau,Joao A. Paulo,Wei Zhang,Tim Ahfeldt,Jiuchun Zhang,Erin F. Cohn,Zhonggang Hou,Jin-Mi Heo,Lee L. Rubin,Sachdev S. Sidhu,Steven P. Gygi,J. Wade Harper +11 more
TL;DR: Temporal digital snapshots of ubiquitin signaling on the mitochondrial outer membrane in embryonic stem cell-derived neurons are developed and the power of this approach to quantify pathway modulators and to mechanistically define the role of PARKIN UBL phosphorylation in pathway activation in induced neurons is demonstrated.
Journal ArticleDOI
Genome-Wide CRISPR-Cas9 Screens Expose Genetic Vulnerabilities and Mechanisms of Temozolomide Sensitivity in Glioblastoma Stem Cells.
Graham MacLeod,Danielle Bozek,Nishani Rajakulendran,Vernon Monteiro,Moloud Ahmadi,Zachary Steinhart,Michelle Kushida,Helen Yu,Fiona J. Coutinho,Florence M.G. Cavalli,Ian Restall,Xiaoguang Hao,Traver Hart,H. Artee Luchman,Samuel Weiss,Peter B. Dirks,Stephane Angers +16 more
TL;DR: Using CRISPR-Cas9 approaches in patient-derived GBM stem cells to interrogate function of the coding genome, actionable pathways responsible for growth are identified, which reveal the gene-essential circuitry of G BM stemness and proliferation.
References
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