1. Biochemistry and Chemical Biology
  2. Cell Biology
Download icon

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
Research Article
  • Cited 75
  • Views 4,688
  • Annotations
Cite this article as: eLife 2016;5:e11794 doi: 10.7554/eLife.11794

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

  • 4,688
    Page views
  • 1,434
    Downloads
  • 75
    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)

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

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

Further reading

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Mikel Garcia-Marcos
    Research Article Updated

    It has become evident that activation of heterotrimeric G-proteins by cytoplasmic proteins that are not G-protein-coupled receptors (GPCRs) plays a role in physiology and disease. Despite sharing the same biochemical guanine nucleotide exchange factor (GEF) activity as GPCRs in vitro, the mechanisms by which these cytoplasmic proteins trigger G-protein-dependent signaling in cells have not been elucidated. Heterotrimeric G-proteins can give rise to two active signaling species, Gα-GTP and dissociated Gβγ, with different downstream effectors, but how non-receptor GEFs affect the levels of these two species in cells is not known. Here, a systematic comparison of GPCRs and three unrelated non-receptor proteins with GEF activity in vitro (GIV/Girdin, AGS1/Dexras1, and Ric-8A) revealed high divergence in their contribution to generating Gα-GTP and free Gβγ in cells directly measured with live-cell biosensors. These findings demonstrate fundamental differences in how receptor and non-receptor G-protein activators promote signaling in cells despite sharing similar biochemical activities in vitro.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Zachary F Mandell et al.
    Research Article

    NusA and NusG are transcription factors that stimulate RNA polymerase pausing in Bacillus subtilis. While NusA was known to function as an intrinsic termination factor in B. subtilis, the role of NusG in this process was unknown. To examine the individual and combinatorial roles that NusA and NusG play in intrinsic termination, Term-seq was conducted in wild type, NusA depletion, DnusG, and NusA depletion DnusG strains. We determined that NusG functions as an intrinsic termination factor that works alone and cooperatively with NusA to facilitate termination at 88% of the 1400 identified intrinsic terminators. Our results indicate that NusG stimulates a sequence-specific pause that assists in the completion of suboptimal terminator hairpins with weak terminal A-U and G-U base pairs at the bottom of the stem. Loss of NusA and NusG leads to global misregulation of gene expression and loss of NusG results in flagella and swimming motility defects.