1. Cell Biology
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Distinct Regulatory Ribosomal Ubiquitylation events are reversible and hierarchically organized

  1. Danielle M Garshott
  2. Elayanambi Sundaramoorthy
  3. Marilyn Leonard
  4. Eric J Bennett  Is a corresponding author
  1. University of California, San Diego, United States
Research Article
  • Cited 7
  • Views 1,524
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Cite this article as: eLife 2020;9:e54023 doi: 10.7554/eLife.54023

Abstract

Activation of the integrated stress response (ISR) or the ribosome-associated quality control (RQC) pathway stimulates regulatory ribosomal ubiquitylation (RRub) on distinct 40S ribosomal proteins, yet the cellular role and fate of ubiquitylated proteins remain unclear. We demonstrate that uS10 and uS5 ubiquitylation are dependent upon eS10 or uS3 ubiquitylation, respectively, suggesting that a hierarchical relationship exists among RRub events establishing a ubiquitin code on ribosomes. We show that stress dependent RRub events diminish after initial stimuli and that RRub loss is the result of demodification by deubiquitylating enzymes rather than protein degradation. Utilizing an optical RQC reporter we identify OTUD3 and USP21 as deubiquitylating enzymes that antagonize ZNF598-mediated 40S ubiquitylation and can limit RQC activation. Critically, cells lacking USP21 or OTUD3 have altered RQC activity and delayed eS10 deubiquitylation indicating a functional role for deubiquitylating enzymes within the RQC pathway.

Article and author information

Author details

  1. Danielle M Garshott

    Section of Cell and Developmental Biology, University of California, San Diego, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4357-1781
  2. Elayanambi Sundaramoorthy

    Section of Cell and Developmental Biology, University of California, San Diego, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1256-9758
  3. Marilyn Leonard

    Section of Cell and Developmental Biology, University of California, San Diego, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Eric J Bennett

    Section of Cell and Developmental Biology, University of California, San Diego, La Jolla, United States
    For correspondence
    e1bennett@ucsd.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1201-3314

Funding

National Institutes of Health (DP2-GM119132)

  • Eric J Bennett

National Institutes of Health (PGM085764)

  • Eric J Bennett

National Science Foundation (DGE-1650112)

  • Danielle M Garshott

The authors declare that the funders had no role in designing this study, data collection, or interpretation.

Reviewing Editor

  1. Rachel Green, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, United States

Publication history

  1. Received: November 28, 2019
  2. Accepted: February 1, 2020
  3. Accepted Manuscript published: February 3, 2020 (version 1)
  4. Version of Record published: March 10, 2020 (version 2)

Copyright

© 2020, Garshott 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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