1. Biochemistry and Chemical Biology
  2. Cell Biology
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The multi-subunit GID/CTLH E3 ligase promotes proliferation and targets the transcription factor Hbp1 for degradation

  1. Fabienne Lampert  Is a corresponding author
  2. Diana Stafa
  3. Algera Goga
  4. Martin Varis Soste
  5. Samuel Gilberto
  6. Natacha Olieric
  7. Paola Picotti
  8. Markus Stoffel
  9. Matthias Peter  Is a corresponding author
  1. ETH Zürich, Switzerland
  2. Paul Scherrer Institute, Switzerland
Research Article
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Cite this article as: eLife 2018;7:e35528 doi: 10.7554/eLife.35528

Abstract

In yeast, the glucose-induced degradation-deficient (GID) E3 ligase selectively degrades superfluous gluconeogenic enzymes. Here we identified all subunits of the mammalian GID/CTLH complex and provide a comprehensive map of its hierarchical organization and step-wise assembly. Biochemical reconstitution demonstrates that the mammalian complex possesses inherent E3 ubiquitin ligase activity, using Ube2H as its cognate E2. Deletions of multiple GID subunits compromise cell proliferation, and this defect is accompanied by deregulation of critical cell cycle markers such as the retinoblastoma (Rb) tumor suppressor, phospho-Histone H3 and Cyclin A. We identify the negative regulator of pro-proliferative genes Hbp1 as a bonafide GID/CTLH proteolytic substrate. Indeed, Hbp1 accumulates in cells lacking GID/CTLH activity, and Hbp1 physically interacts and is ubiquitinated in vitro by reconstituted GID/CTLH complexes. Our biochemical and cellular analysis thus demonstrates that the GID/CTLH complex prevents cell cycle exit in G1, at least in part by degrading Hbp1.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files contain the complete lists of mass spectrometry results and SAINT scores

Article and author information

Author details

  1. Fabienne Lampert

    Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
    For correspondence
    fabienne.lampert@bc.biol.ethz.ch
    Competing interests
    The authors declare that no competing interests exist.

  2. Diana Stafa

    Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  3. Algera Goga

    Institute of Molecular Health Sciences, ETH Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  4. Martin Varis Soste

    Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  5. Samuel Gilberto

    Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  6. Natacha Olieric

    Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  7. Paola Picotti

    Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  8. Markus Stoffel

    Institute of Molecular Health Sciences, ETH Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  9. Matthias Peter

    Institute of Biochemistry, ETH Zürich, Zürich, Switzerland
    For correspondence
    matthias.peter@bc.biol.ethz.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2160-6824

Funding

Swiss National Science Foundation

  • Markus Stoffel
  • Matthias Peter

Human Frontier Science Program (LT- 000376/2014-L)

  • Fabienne Lampert

European Research Council

  • Matthias Peter

ETH Zurich

  • Paola Picotti
  • Markus Stoffel
  • Matthias Peter

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

Ethics

Animal experimentation: All animal experiments were approved by the Kantonale Veterinäramt Zürich.

Reviewing Editor

  1. Ivan Dikic, Goethe University Frankfurt, Germany

Publication history

  1. Received: January 30, 2018
  2. Accepted: June 16, 2018
  3. Accepted Manuscript published: June 18, 2018 (version 1)
  4. Version of Record published: July 9, 2018 (version 2)

Copyright

© 2018, Lampert 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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