1. System-Wide Modulation of HECT E3 Ligases with Selective Ubiquitin Variant Probes
- Author
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Emmanuel Lemichez, Brenda A. Schulman, Zhenyue Hao, Sachdev S. Sidhu, Nan Li, Ryan Murchie, Wei Zhang, Maria A. Sartori, Kuen-Phon Wu, Jason Moffat, John R. Walker, Avinash Persaud, Yi Sheng, Alban Ordureau, Jicheng Hu, Chong Jiang, Manjeet Mukherjee, Kevin R. Brown, Yufeng Tong, Yanjun Li, Junjie Chen, Anne Doye, Peter Y. Mercredi, Daniela Rotin, Hari B. Kamadurai, J. Wade Harper, Donnelly Center Cellular and Biomolecular Research [Univ. Toronto], University of Toronto, St Jude Children's Research Hospital, Department of Cell Biology, Harvard Medical School [Boston] (HMS), Program in Cell Biology [Toronto], The Hospital for sick children [Toronto] (SickKids)-Department of Biochemistry [University of Toronto], University of Toronto-University of Toronto, Structural Genomics Consortium, The University of Texas M.D. Anderson Cancer Center [Houston], Centre méditerranéen de médecine moléculaire (C3M), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM), University of York [York, UK], Campbell Family Institute for Breast Cancer Research, University Health Network, Department of Pharmacology and Toxicology [Toronto], Department of Molecular Genetics [Toronto], The Donnelly Centre for Cellular and Biomolecular Research [Toronto, ON, Canada] (CCBR), J.W.H. was supported by NIH (R37NS083524 and GM095567). A.O. was supported by an Edward R. and Anne G. Lefler Center postdoctoral fellowship. J.W.H. is a consultant for Millennium: the Takada Oncology Company and Biogen. D.R. holds a Canada Research Chair (Tier 1 in Biochemistry and Signal Transduction) and was supported by CIHR (MOP#130422). B.A.S. is an investigator of the Howard Hughes Medical Institute (HHMI) and was supported by ALSAC, NIH R37GM065930 and P30CA021765. NECAT and APS were supported by NIH P41 GM103403 and DOE Contract DE-AC02-06CH11357. W.Z. was supported by a CIHR postdoctoral fellowship. S.S.S. and J.M. were supported by CIHR (MOP#111149 and 136956). The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada, Innovative Medicines Initiative (EU/EFPIA) (ULTRA-DD grant no. 115766), Janssen, Merck & Co., Novartis Pharma AG, Ontario Ministry of Economic Development and Innovation, Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and the Wellcome Trust., and We thank members of the Sidhu and Schulman groups for helpful comments. We thank Andrew Vorobyov, Eva Chou, Yogesh Hooda, Jun Gu, and Aiping Dong for technical assistance. We are indebted to Pankaj Garg, Andreas Ernst, Abiodun Ogunjimi, Clare Jeon, Satra Nim, Frank Sicheri, and Hongrui Wang for reagents and advice.
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Models, Molecular ,0301 basic medicine ,Subfamily ,[SDV]Life Sciences [q-bio] ,MESH: Catalytic Domain ,NEDD4 ,[SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC] ,MESH: Ubiquitin/chemistry ,MESH: Ubiquitin/metabolism ,Madin Darby Canine Kidney Cells ,MESH: Dogs ,Ubiquitin ,Cell Movement ,Catalytic Domain ,[SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB] ,MESH: Animals ,MESH: Cell Movement ,chemistry.chemical_classification ,MESH: Organoids/metabolism ,biology ,MESH: Ubiquitin-Protein Ligases/chemistry ,Cell migration ,Cell biology ,MESH: Ubiquitin/genetics ,Organoids ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,MESH: Models, Molecular ,MESH: HCT116 Cells ,Ubiquitin-Protein Ligases ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,macromolecular substances ,Cell Line ,03 medical and health sciences ,Dogs ,Peptide Library ,Animals ,Humans ,Molecular Biology ,NEDD4L ,DNA ligase ,MESH: Humans ,MESH: Organoids/cytology ,MESH: Madin Darby Canine Kidney Cells ,MESH: Ubiquitin-Protein Ligases/metabolism ,Cell Biology ,HCT116 Cells ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,MESH: Cell Line ,030104 developmental biology ,chemistry ,biology.protein ,MESH: Peptide Library ,Genetic screen - Abstract
Comment in : A Billion Ubiquitin Variants to Probe and Modulate the UPS. [Mol Cell. 2016]; International audience; HECT-family E3 ligases ubiquitinate protein substrates to control virtually every eukaryotic process and are misregulated in numerous diseases. Nonetheless, understanding of HECT E3s is limited by a paucity of selective and potent modulators. To overcome this challenge, we systematically developed ubiquitin variants (UbVs) that inhibit or activate HECT E3s. Structural analysis of 6 HECT-UbV complexes revealed UbV inhibitors hijacking the E2-binding site and activators occupying a ubiquitin-binding exosite. Furthermore, UbVs unearthed distinct regulation mechanisms among NEDD4 subfamily HECTs and proved useful for modulating therapeutically relevant targets of HECT E3s in cells and intestinal organoids, and in a genetic screen that identified a role for NEDD4L in regulating cell migration. Our 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.
- Published
- 2016