1. Cancer Biology
  2. Cell Biology

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
https://doi.org/10.7554/eLife.77780
Share this article
Cite this article
  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
  2. Download BibTeX
  3. Download .RIS

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.

Metrics

  • 2,687
    views
  • 753
    downloads
  • 3
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  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

Share this article

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

Categories and tags

Research organism

Further reading

    1. Cancer Biology

    Recording and classifying MET receptor mutations in cancers

    Célia Guérin, David Tulasne
    Review Article

    Tyrosine kinase inhibitors (TKI) directed against MET have been recently approved to treat advanced non-small cell lung cancer (NSCLC) harbouring activating MET mutations. This success is the consequence of a long characterization of MET mutations in cancers, which we propose to outline in this review. MET, a receptor tyrosine kinase (RTK), displays in a broad panel of cancers many deregulations liable to promote tumour progression. The first MET mutation was discovered in 1997, in hereditary papillary renal cancer (HPRC), providing the first direct link between MET mutations and cancer development. As in other RTKs, these mutations are located in the kinase domain, leading in most cases to ligand-independent MET activation. In 2014, novel MET mutations were identified in several advanced cancers, including lung cancers. These mutations alter splice sites of exon 14, causing in-frame exon 14 skipping and deletion of a regulatory domain. Because these mutations are not located in the kinase domain, they are original and their mode of action has yet to be fully elucidated. Less than five years after the discovery of such mutations, the efficacy of a MET TKI was evidenced in NSCLC patients displaying MET exon 14 skipping. Yet its use led to a resistance mechanism involving acquisition of novel and already characterized MET mutations. Furthermore, novel somatic MET mutations are constantly being discovered. The challenge is no longer to identify them but to characterize them in order to predict their transforming activity and their sensitivity or resistance to MET TKIs, in order to adapt treatment.

    1. Cancer Biology
    2. Genetics and Genomics

    Convergent epigenetic evolution drives relapse in acute myeloid leukemia

    Kevin Nuno, Armon Azizi ... Ravindra Majeti
    Research Article

    Relapse of acute myeloid leukemia (AML) is highly aggressive and often treatment refractory. We analyzed previously published AML relapse cohorts and found that 40% of relapses occur without changes in driver mutations, suggesting that non-genetic mechanisms drive relapse in a large proportion of cases. We therefore characterized epigenetic patterns of AML relapse using 26 matched diagnosis-relapse samples with ATAC-seq. This analysis identified a relapse-specific chromatin accessibility signature for mutationally stable AML, suggesting that AML undergoes epigenetic evolution at relapse independent of mutational changes. Analysis of leukemia stem cell (LSC) chromatin changes at relapse indicated that this leukemic compartment underwent significantly less epigenetic evolution than non-LSCs, while epigenetic changes in non-LSCs reflected overall evolution of the bulk leukemia. Finally, we used single-cell ATAC-seq paired with mitochondrial sequencing (mtscATAC) to map clones from diagnosis into relapse along with their epigenetic features. We found that distinct mitochondrially-defined clones exhibit more similar chromatin accessibility at relapse relative to diagnosis, demonstrating convergent epigenetic evolution in relapsed AML. These results demonstrate that epigenetic evolution is a feature of relapsed AML and that convergent epigenetic evolution can occur following treatment with induction chemotherapy.

    1. Cancer Biology
    2. Cell Biology

    Early recovery of proteasome activity in cells pulse-treated with proteasome inhibitors is independent of DDI2

    Ibtisam Ibtisam, Alexei F Kisselev
    Short Report

    Rapid recovery of proteasome activity may contribute to intrinsic and acquired resistance to FDA-approved proteasome inhibitors. Previous studies have demonstrated that the expression of proteasome genes in cells treated with sub-lethal concentrations of proteasome inhibitors is upregulated by the transcription factor Nrf1 (NFE2L1), which is activated by a DDI2 protease. Here, we demonstrate that the recovery of proteasome activity is DDI2-independent and occurs before transcription of proteasomal genes is upregulated but requires protein translation. Thus, mammalian cells possess an additional DDI2 and transcription-independent pathway for the rapid recovery of proteasome activity after proteasome inhibition.