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Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle

Mosadeghi, Ruzbeh and Reichermeier, Kurt M. and Winkler, Martin and Schreiber, Anne and Reitsma, Justin M. and Zhang, Yaru and Stengel, Florian and Cao, Junyue and Kim, Minsoo and Sweredoski, Michael J. and Hess, Sonja and Leitner, Alexander and Aebersold, Ruedi and Peter, Matthias and Deshaies, Raymond J. and Enchev, Radoslav I. (2016) Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle. eLife, 5 . Art. No. e12102. ISSN 2050-084X. PMCID PMC4878873.

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The COP9-Signalosome (CSN) regulates cullin–RING ubiquitin ligase (CRL) activity and assembly by cleaving Nedd8 from cullins. Free CSN is autoinhibited, and it remains unclear how it becomes activated. We combine structural and kinetic analyses to identify mechanisms that contribute to CSN activation and Nedd8 deconjugation. Both CSN and neddylated substrate undergo large conformational changes upon binding, with important roles played by the N-terminal domains of Csn2 and Csn4 and the RING domain of Rbx1 in enabling formation of a high affinity, fully active complex. The RING domain is crucial for deneddylation, and works in part through conformational changes involving insert-2 of Csn6. Nedd8 deconjugation and re-engagement of the active site zinc by the autoinhibitory Csn5 glutamate-104 diminish affinity for Cul1/Rbx1 by ~100-fold, resulting in its rapid ejection from the active site. Together, these mechanisms enable a dynamic deneddylation-disassembly cycle that promotes rapid remodeling of the cellular CRL network.

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URLURL TypeDescription CentralArticle Paper
Sweredoski, Michael J.0000-0003-0878-3831
Hess, Sonja0000-0002-5904-9816
Deshaies, Raymond J.0000-0002-3671-9354
Additional Information:© 2016, Mosadeghi et al. This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited. Received October 5, 2015. Accepted March 30, 2016. Published March 31, 2016. We thank B Schulman for Skp1/Skp2 protein as well as for expression plasmids and E Morris for thin carbon for EM grids. We thank A Bernini and A Ragheb for technical assistance with protein work, P Tittmann for technical support at the ScopeM facility, members of the Ban lab for advice and Annie Moradian and Roxana Eggleston-Rangel for mass spectrometry support at PEL. We also are grateful to SO Shan, E Morris and T Stuwe for advice and A Smith, SO Shan, and D Barford for comments on the manuscript. RM was supported by a Lee-Ramo Life Sciences Fellowship, RIE was supported by an ETH Pioneer, a Marie Curie and an EMBO short-term fellowship, and AS by a Marie Curie fellowship. FS acknowledges funding from the Wellcome Trust (Grant 095951) and the German Science Foundation Collaborative Research Center (SFB) 969. The Peter laboratory is funded by an ERC advanced grant, the SNF and ETHZ, and the Aebersold laboratory is supported by ETH Zurich, and an ERC advanced grant. This work was supported in part by NIH CA164803 to RJD RJD is an Investigator of and was supported by the Howard Hughes Medical Institute. MJS and SH were supported by the Gordon and Betty Moore Foundation, through Grant GBMF775 and the Beckman Institute. JMR was supported by National Institute of General Medical Sciences award F32GM112308. Competing interests: Raymond J Deshaies, Reviewing editor, eLife. The other authors declare that no competing interests exist.
Funding AgencyGrant Number
European Molecular Biology Organization (EMBO)UNSPECIFIED
Marie Curie FellowshipUNSPECIFIED
Wellcome Trust095951
Deutsche Forschungsgemeinschaft (DFG)SFB 969
European Research Council (ERC)UNSPECIFIED
Swiss National Fund (SNF)UNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Gordon and Betty Moore FoundationGBMF775
Caltech Beckman InstituteUNSPECIFIED
PubMed Central ID:PMC4878873
Record Number:CaltechAUTHORS:20160405-075712117
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Official Citation:Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle Ruzbeh Mosadeghi Kurt M Reichermeier Martin Winkler Anne Schreiber Justin M Reitsma Yaru Zhang Florian Stengel Junyue Cao Minsoo Kim Michael J Sweredoski Sonja Hess Alexander Leitner Ruedi Aebersold Matthias Peter Raymond J Deshaies Radoslav I Enchev eLife 2016;10.7554/eLife.12102
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65914
Deposited By: Ruth Sustaita
Deposited On:06 Apr 2016 21:36
Last Modified:03 Oct 2019 09:51

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