The integrated stress response remodels the microtubule organizing center to clear unfolded proteins following proteotoxic stress

  1. Brian Hurwitz
  2. Nicola Guzzi
  3. Anita Gola
  4. Vincent F Fiore
  5. Ataman Sendoel
  6. Maria Nikolova
  7. Douglas Barrows
  8. Thomas S Carroll
  9. H Amalia Pasolli
  10. Elaine Fuchs  Is a corresponding author
  1. Howard Hughes Medical Institute, The Rockefeller University, United States
  2. University of Zurich, Switzerland
  3. Rockefeller University, United States

Abstract

Cells encountering stressful situations activate the integrated stress response (ISR) pathway to limit protein synthesis and redirect translation to better cope. The ISR has also been implicated in cancers, but redundancies in the stress-sensing kinases that trigger the ISR have posed hurdles to dissecting physiological relevance. To overcome this challenge, we targeted the regulatory node of these kinases, namely the S51 phosphorylation site of eukaryotic translation initiation factor eIF2α and genetically replaced eIF2α with eIF2α-S51A in mouse squamous cell carcinoma (SCC) stem cells of skin. While inconsequential under normal growth conditions, the vulnerability of this ISR-null state was unveiled when SCC stem cells experienced proteotoxic stress. Seeking mechanistic insights into the protective roles of the ISR, we combined ribosome profiling and functional approaches to identify and probe the functional importance of translational differences between ISR-competent and ISR-null SCC stem cells when exposed to proteotoxic stress. In doing so, we learned that the ISR redirects translation to centrosomal proteins that orchestrate the microtubule dynamics needed to efficiently concentrate unfolded proteins at the microtubule organizing center so that they can be cleared by the perinuclear degradation machinery. Thus, rather than merely maintaining survival during proteotoxic stress, the ISR also functions in promoting cellular recovery once the stress has subsided. Remarkably, this molecular program is unique to transformed skin stem cells hence exposing a vulnerability in cancer that could be exploited therapeutically.

Data availability

Sequencing data have been deposited in GEO under accession codes GSE193945

The following data sets were generated

Article and author information

Author details

  1. Brian Hurwitz

    Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  2. Nicola Guzzi

    Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3898-8064
  3. Anita Gola

    Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  4. Vincent F Fiore

    Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  5. Ataman Sendoel

    Institute for Regenerative Medicine, University of Zurich, Zurich, Switzerland
    Competing interests
    No competing interests declared.
  6. Maria Nikolova

    Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  7. Douglas Barrows

    Bioinformatics Resource Center, Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  8. Thomas S Carroll

    Bioinformatics Resouce Center, Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  9. H Amalia Pasolli

    Electron Microscopy Resource Center, Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  10. Elaine Fuchs

    Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    For correspondence
    fuchs@rockefeller.edu
    Competing interests
    Elaine Fuchs, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0978-5137

Funding

Howard Hughes Medical Institute

  • Elaine Fuchs

Ruth Kirschstein NIH Predoctoral Fellow (F30CA236239)

  • Brian Hurwitz

Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional Medical Scientist Training Program (T32GM007739)

  • Brian Hurwitz

HHMI Jane Coffin Childs Associate

  • Nicola Guzzi

Damon Runyon Cancer Research Foundation National Mah Jongg League Fellowship (DRG 2409-20)

  • Anita Gola

National Institutes of Health (R01-AR27883)

  • Elaine Fuchs

Robertson Foundation

  • Brian Hurwitz

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

Reviewing Editor

  1. Marianne E Bronner, California Institute of Technology, United States

Ethics

Animal experimentation: Animal experimentation: All animal procedures used in this study are described in our #20066H protocol named Development and Differentiation in the Skin, which had been previously reviewed and approved by the Rockefeller University Institutional Animal Care and Use Committee (IACUC).

Version history

  1. Received: February 10, 2022
  2. Preprint posted: February 14, 2022 (view preprint)
  3. Accepted: June 27, 2022
  4. Accepted Manuscript published: June 27, 2022 (version 1)
  5. Version of Record published: July 20, 2022 (version 2)

Copyright

© 2022, Hurwitz 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. Brian Hurwitz
  2. Nicola Guzzi
  3. Anita Gola
  4. Vincent F Fiore
  5. Ataman Sendoel
  6. Maria Nikolova
  7. Douglas Barrows
  8. Thomas S Carroll
  9. H Amalia Pasolli
  10. Elaine Fuchs
(2022)
The integrated stress response remodels the microtubule organizing center to clear unfolded proteins following proteotoxic stress
eLife 11:e77780.
https://doi.org/10.7554/eLife.77780

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https://doi.org/10.7554/eLife.77780