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The steady-state repertoire of human SCF Ubiquitin ligase complexes does not require ongoing Nedd8 conjugation

Lee, J. Eugene and Sweredoski, Michael J. and Graham, Robert L. J. and Kolawa, Natalie J. and Smith, Geoffrey T. and Hess, Sonja and Deshaies, Raymond J. (2011) The steady-state repertoire of human SCF Ubiquitin ligase complexes does not require ongoing Nedd8 conjugation. Molecular and Cellular Proteomics, 10 (5). Art. No. M110.006460. ISSN 1535-9476. PMCID PMC3098594. https://resolver.caltech.edu/CaltechAUTHORS:20110526-100158794

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Abstract

The human genome encodes 69 different F-box proteins (FBPs), each of which can potentially assemble with Skp1-Cul1-RING to serve as the substrate specificity subunit of an SCF ubiquitin ligase complex. SCF activity is switched on by conjugation of the ubiquitin- like protein Nedd8 to Cul1. Cycles of Nedd8 conjugation and deconjugation acting in conjunction with the Cul1-sequestering factor Cand1 are thought to control dynamic cycles of SCF assembly and disassembly, which would enable a dynamic equilibrium between the Cul1- RING catalytic core of SCF and the cellular repertoire of FBPs. To test this hypothesis, we determined the cellular composition of SCF complexes and evaluated the impact of Nedd8 conjugation on this steady-state. At least 42 FBPs assembled with Cul1 in HEK 293 cells, and the levels of Cul1-bound FBPs varied by over two orders of magnitude. Unexpectedly, quantitative mass spectrometry revealed that blockade of Nedd8 conjugation led to a modest increase, rather than a decrease, in the overall level of most SCF complexes. We suggest that multiple mechanisms including FBP dissociation and turnover cooperate to maintain the cellular pool of SCF ubiquitin ligases.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1074/mcp.M110.006460 DOIArticle
http://www.mcponline.org/content/10/5/M110.006460PublisherArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3098594/PubMed CentralArticle
ORCID:
AuthorORCID
Sweredoski, Michael J.0000-0003-0878-3831
Hess, Sonja0000-0002-5904-9816
Deshaies, Raymond J.0000-0002-3671-9354
Additional Information:© 2011 The American Society for Biochemistry and Molecular Biology, Inc. Received November 16, 2010. Accepted December 17, 2010. J. E. Lee was supported by the Ruth L. Kirschstein NRSA fellowship (CA138126) and the Proteome Exploration lab was supported by the Beckman Institute at Caltech and an award from the Gordon and Betty Moore Foundation. R. J. D. is an Investigator of the Howard Hughes Medical Institute and this work was supported in part by HHMI and an NIH grant (GM065997) to R. J. D. Acknowledgments—We thank P. Kaiser for the HTBH tag and J. Wade Harper for generously sharing results prior to publication. MLN4924 was a generous gift from Millennium: The Takeda Oncology Company.
Funders:
Funding AgencyGrant Number
NIH Predoctoral FellowshipCA138126
Caltech Beckman InstituteUNSPECIFIED
Gordon and Betty Moore FoundationUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NIHGM065997
Issue or Number:5
PubMed Central ID:PMC3098594
Record Number:CaltechAUTHORS:20110526-100158794
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20110526-100158794
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:23807
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:17 Jun 2011 18:04
Last Modified:03 Oct 2019 02:50

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