The Rqc2/Tae2 subunit of the Ribosome-Associated Quality Control (RQC) complex marks ribosome-stalled nascent polypeptide chains for aggregation

  1. Ryo Yonashiro
  2. Erich B Tahara
  3. Mario H Bengtson
  4. Maria Khokhrina
  5. Holger Lorenz
  6. Kai-Chun Chen
  7. Yu Kigoshi-Tansho
  8. Jeffrey N Savas
  9. John R Yates
  10. Steve A Kay
  11. Elizabeth A Craig
  12. Axel Mogk
  13. Bernd Bukau
  14. Claudio AP Joazeiro  Is a corresponding author
  1. The Scripps Research Institute, United States
  2. University of São Paulo, Brazil
  3. University of Campinas, Brazil
  4. Zentrum für Molekulare Biologie der Universität Heidelberg, Germany
  5. Northwestern University, United States
  6. University of Wisconsin - Madison, United States

Abstract

Ribosome stalling during translation can be harmful, and is surveyed by a conserved quality control pathway that targets the associated mRNA and nascent polypeptide chain (NC). In this pathway, the ribosome-associated quality control (RQC) complex promotes the ubiquitylation and degradation of NCs remaining stalled in the 60S subunit. NC stalling is recognized by the Rqc2/Tae2 RQC subunit, which also stabilizes binding of the E3 ligase, Listerin/Ltn1. Additionally, Rqc2 modifies stalled NCs with a carboxy-terminal, Ala- and Thr-containing extension-the 'CAT tail.' However, the function of CAT tails and fate of CAT tail-modified ('CATylated') NCs has remained unknown. Here we show that CATylation mediates NC aggregation. NC CATylation and aggregation could be observed by inactivating Ltn1 or by analyzing NCs with limited ubiquitylation potential, suggesting that inefficient targeting by Ltn1 favors the Rqc2-mediated reaction. These findings uncover a translational stalling-dependent protein aggregation mechanism, and provide evidence that proteins can become marked for aggregation.

Article and author information

Author details

  1. Ryo Yonashiro

    Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Erich B Tahara

    University of São Paulo, São Paulo, Brazil
    Competing interests
    The authors declare that no competing interests exist.
  3. Mario H Bengtson

    University of Campinas, São Paulo, Brazil
    Competing interests
    The authors declare that no competing interests exist.
  4. Maria Khokhrina

    Deutsches Krebsforschungszentrum, DKFZ-ZMBH Alliance, Zentrum für Molekulare Biologie der Universität Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Holger Lorenz

    Deutsches Krebsforschungszentrum, DKFZ-ZMBH Alliance, Zentrum für Molekulare Biologie der Universität Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Kai-Chun Chen

    Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Yu Kigoshi-Tansho

    Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Jeffrey N Savas

    Department of Neurology, Feinberg School of Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. John R Yates

    Department of Chemical Physiology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Steve A Kay

    Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Elizabeth A Craig

    Department of Biochemistry, University of Wisconsin - Madison, Wisconsin, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Axel Mogk

    Deutsches Krebsforschungszentrum, DKFZ-ZMBH Alliance, Zentrum für Molekulare Biologie der Universität Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  13. Bernd Bukau

    Deutsches Krebsforschungszentrum, DKFZ-ZMBH Alliance, Zentrum für Molekulare Biologie der Universität Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  14. Claudio AP Joazeiro

    Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, United States
    For correspondence
    joazeiro@scripps.edu
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Ivan Dikic, Goethe University Medical School, Germany

Publication history

  1. Received: September 23, 2015
  2. Accepted: March 3, 2016
  3. Accepted Manuscript published: March 4, 2016 (version 1)
  4. Version of Record published: March 14, 2016 (version 2)

Copyright

© 2016, Yonashiro 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

  • 5,147
    Page views
  • 1,498
    Downloads
  • 91
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

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. Ryo Yonashiro
  2. Erich B Tahara
  3. Mario H Bengtson
  4. Maria Khokhrina
  5. Holger Lorenz
  6. Kai-Chun Chen
  7. Yu Kigoshi-Tansho
  8. Jeffrey N Savas
  9. John R Yates
  10. Steve A Kay
  11. Elizabeth A Craig
  12. Axel Mogk
  13. Bernd Bukau
  14. Claudio AP Joazeiro
(2016)
The Rqc2/Tae2 subunit of the Ribosome-Associated Quality Control (RQC) complex marks ribosome-stalled nascent polypeptide chains for aggregation
eLife 5:e11794.
https://doi.org/10.7554/eLife.11794
  1. Further reading

Further reading

    1. Biochemistry and Chemical Biology
    2. Neuroscience
    Jinli Geng, Yingjun Tang ... Xiaodong Liu
    Research Article Updated

    Dynamic Ca2+ signals reflect acute changes in membrane excitability, and also mediate signaling cascades in chronic processes. In both cases, chronic Ca2+ imaging is often desired, but challenged by the cytotoxicity intrinsic to calmodulin (CaM)-based GCaMP, a series of genetically-encoded Ca2+ indicators that have been widely applied. Here, we demonstrate the performance of GCaMP-X in chronic Ca2+ imaging of cortical neurons, where GCaMP-X by design is to eliminate the unwanted interactions between the conventional GCaMP and endogenous (apo)CaM-binding proteins. By expressing in adult mice at high levels over an extended time frame, GCaMP-X showed less damage and improved performance in two-photon imaging of sensory (whisker-deflection) responses or spontaneous Ca2+ fluctuations, in comparison with GCaMP. Chronic Ca2+ imaging of one month or longer was conducted for cultured cortical neurons expressing GCaMP-X, unveiling that spontaneous/local Ca2+ transients progressively developed into autonomous/global Ca2+ oscillations. Along with the morphological indices of neurite length and soma size, the major metrics of oscillatory Ca2+, including rate, amplitude and synchrony were also examined. Dysregulations of both neuritogenesis and Ca2+ oscillations became discernible around 2–3 weeks after virus injection or drug induction to express GCaMP in newborn or mature neurons, which were exacerbated by stronger or prolonged expression of GCaMP. In contrast, neurons expressing GCaMP-X were significantly less damaged or perturbed, altogether highlighting the unique importance of oscillatory Ca2+ to neural development and neuronal health. In summary, GCaMP-X provides a viable solution for Ca2+ imaging applications involving long-time and/or high-level expression of Ca2+ probes.

    1. Biochemistry and Chemical Biology
    2. Chromosomes and Gene Expression
    Radhika A Varier, Theodora Sideri ... Folkert Jacobus van Werven
    Research Article

    N6-methyladenosine (m6A) RNA modification impacts mRNA fate primarily via reader proteins, which dictate processes in development, stress, and disease. Yet little is known about m6A function in Saccharomyces cerevisiae, which occurs solely during early meiosis. Here we perform a multifaceted analysis of the m6A reader protein Pho92/Mrb1. Cross-linking immunoprecipitation analysis reveals that Pho92 associates with the 3’end of meiotic mRNAs in both an m6A-dependent and independent manner. Within cells, Pho92 transitions from the nucleus to the cytoplasm, and associates with translating ribosomes. In the nucleus Pho92 associates with target loci through its interaction with transcriptional elongator Paf1C. Functionally, we show that Pho92 promotes and links protein synthesis to mRNA decay. As such, the Pho92-mediated m6A-mRNA decay is contingent on active translation and the CCR4-NOT complex. We propose that the m6A reader Pho92 is loaded co-transcriptionally to facilitate protein synthesis and subsequent decay of m6A modified transcripts, and thereby promotes meiosis.