Cyclin F drives proliferation through SCF-dependent degradation of the retinoblastoma-like tumor suppressor p130/RBL2

  1. Taylor P Enrico
  2. Wayne Stallaert
  3. Elizaveta T Wick
  4. Peter Ngoi
  5. Xianxi Wang
  6. Seth M Rubin
  7. Nicholas G Brown
  8. Jeremy Purvis
  9. Michael J Emanuele  Is a corresponding author
  1. University of North Carolina at Chapel Hill, United States
  2. University of California, Santa Cruz, United States

Abstract

Cell cycle gene expression programs fuel proliferation and are universally dysregulated in cancer. The retinoblastoma (RB)-family of proteins, RB1, RBL1/p107 and RBL2/p130, coordinately repress cell cycle gene expression, inhibiting proliferation and suppressing tumorigenesis. Phosphorylation of RB-family proteins by cyclin dependent kinases is firmly established. Like phosphorylation, ubiquitination is essential to cell cycle control, and numerous proliferative regulators, tumor suppressors, and oncoproteins are ubiquitinated. However, little is known about the role of ubiquitin signaling in controlling RB-family proteins. A systems genetics analysis of CRISPR/Cas9 screens suggested the potential regulation of the RB-network by cyclin F, a substrate recognition receptor for the SCF family of E3 ligases. We demonstrate that RBL2/p130 is a direct substrate of SCFcyclin F. We map a cyclin F regulatory site to a flexible linker in the p130 pocket domain, and show that this site mediates binding, stability, and ubiquitination. Expression of a mutant version of p130, which cannot be ubiquitinated, severely impaired proliferative capacity and cell cycle progression. Consistently, we observed reduced expression of cell cycle gene transcripts, as well a reduced abundance of cell cycle proteins, analyzed by quantitative, iterative immunofluorescent imaging. These data suggest a key role for SCFcyclin F in the CDK-RB network and raise the possibility that aberrant p130 degradation could dysregulate the cell cycle in human cancers.

Data availability

Unprocessed, uncropped, immunoblots are made available in the supplemental source data. All raw, unprocessed imaging data is available at Dryad. Raw data related to cell proliferation assays (cell counting and Presto-blue analysis), RT-qPCR, immunoblot quantification for cycloheximide chase experiments and flow cytometry is available in the supplemental source data. All reagents related to this work will be made fully available upon request.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Taylor P Enrico

    Department of Pharmacology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Wayne Stallaert

    Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Elizaveta T Wick

    Department of Pharmacology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Peter Ngoi

    Department of Chemistry and Biochemistry, University of California, Santa Cruz, Santa Cruz, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Xianxi Wang

    Department of Pharmacology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Seth M Rubin

    Department of Chemistry and Biochemistry, University of California, Santa Cruz, Santa Cruz, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1670-4147
  7. Nicholas G Brown

    Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Jeremy Purvis

    Department of Pharmacology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6963-0524
  9. Michael J Emanuele

    Department of Pharmacology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    For correspondence
    emanuele@email.unc.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4104-7449

Funding

National Institute of General Medical Sciences (R01GM120309)

  • Taylor P Enrico
  • Xianxi Wang
  • Michael J Emanuele

National Institute of General Medical Sciences (NIGMS,T32 GM007040)

  • Taylor P Enrico

National Institute of General Medical Sciences (R01GM134231)

  • Taylor P Enrico
  • Xianxi Wang
  • Michael J Emanuele

National Institute of General Medical Sciences (R35GM128855)

  • Elizaveta T Wick
  • Nicholas G Brown

American Cancer Society (RSG-18-220-01-TBG)

  • Taylor P Enrico
  • Xianxi Wang
  • Michael J Emanuele

National Cancer Institute (R01CA163834)

  • Elizaveta T Wick

National Institute of General Medical Sciences (R01GM127707)

  • Peter Ngoi
  • Seth M Rubin

National Institute of General Medical Sciences (GM138834)

  • Wayne Stallaert
  • Jeremy Purvis

National Institute of General Medical Sciences (DP2-HD091800)

  • Wayne Stallaert
  • Jeremy Purvis

National Science Foundation (1845796)

  • Jeremy Purvis

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Silke Hauf, Virginia Tech, United States

Version history

  1. Preprint posted: April 24, 2021 (view preprint)
  2. Received: May 26, 2021
  3. Accepted: November 19, 2021
  4. Accepted Manuscript published: December 1, 2021 (version 1)
  5. Version of Record published: December 14, 2021 (version 2)

Copyright

© 2021, Enrico 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.

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  1. Taylor P Enrico
  2. Wayne Stallaert
  3. Elizaveta T Wick
  4. Peter Ngoi
  5. Xianxi Wang
  6. Seth M Rubin
  7. Nicholas G Brown
  8. Jeremy Purvis
  9. Michael J Emanuele
(2021)
Cyclin F drives proliferation through SCF-dependent degradation of the retinoblastoma-like tumor suppressor p130/RBL2
eLife 10:e70691.
https://doi.org/10.7554/eLife.70691

Share this article

https://doi.org/10.7554/eLife.70691

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