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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Journal ArticleDOI
The linear ubiquitin-specific deubiquitinase gumby regulates angiogenesis
Elena Rivkin,Stephanie M. Almeida,Stephanie M. Almeida,Derek F. Ceccarelli,Yu Chi Juang,Teresa A. MacLean,Teresa A. MacLean,Tharan Srikumar,Hao Huang,Wade H. Dunham,Wade H. Dunham,Ryutaro Fukumura,Gang Xie,Yoichi Gondo,Brian Raught,Anne-Claude Gingras,Anne-Claude Gingras,Frank Sicheri,Frank Sicheri,Sabine P. Cordes,Sabine P. Cordes +20 more
TL;DR: Support is provided for the biological importance of linear (de)ubiquitination in angiogenesis, craniofacial and neural development and in modulating Wnt signalling.
Journal ArticleDOI
Inhibition of 53BP1 favors homology-dependent DNA repair and increases CRISPR-Cas9 genome-editing efficiency.
Marella D. Canny,Nathalie Moatti,Leo C. K. Wan,Leo C. K. Wan,Amélie Fradet-Turcotte,Amélie Fradet-Turcotte,Danielle Krasner,Pedro A. Mateos-Gómez,Michal Zimmermann,Alexandre Orthwein,Alexandre Orthwein,Yu-Chi Juang,Wei Zhang,Sylvie M. Noordermeer,Eduardo Seclén,Marcus D. Wilson,Andrew Vorobyov,Meagan Munro,Andreas Ernst,Andreas Ernst,Timothy F. Ng,Timothy F. Ng,Tiffany Cho,Tiffany Cho,Paula M. Cannon,Sachdev S. Sidhu,Frank Sicheri,Frank Sicheri,Daniel Durocher,Daniel Durocher +29 more
TL;DR: expression of one variant, named i53 (inhibitor of 53BP1), in human and mouse cells, blocked accumulation of 53 BP1 at sites of DNA damage and improved gene targeting and chromosomal gene conversion with either double-stranded DNA or single-strander oligonucleotide donors by up to 5.6-fold.
Journal ArticleDOI
Structural Basis for the Ubiquitin-Linkage Specificity and deISGylating Activity of SARS-CoV Papain-Like Protease
TL;DR: This work proposes that PLpro's higher affinity for K48-linked ubiquitin chains and ISG15 stems from a bivalent mechanism of binding, where two Ubiquitin-like domains prefer to bind in the palm domain of PLpro with the most distal ubiquitIn domain interacting with a “ridge” region of the thumb domain.
Journal ArticleDOI
Regulation of Proteolysis by Human Deubiquitinating Enzymes
Ziad M. Eletr,Keith D. Wilkinson +1 more
TL;DR: The scope and regulation of the ubiquitin pathway is very similar to that of phosphorylation, with the DUBs serving the same functions as the phosphatase.
Journal ArticleDOI
Ubiquitination and the Regulation of Membrane Proteins.
TL;DR: The mechanisms and functions of ubiquitination of membrane proteins are summarized and specific examples of Ubiquitin-dependent regulation of membrane Protein Regulation are provided to downregulate the physiological outcomes.
References
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