1. Biochemistry and Chemical Biology
  2. Cell Biology
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A SPOPL/Cullin-3 ubiquitin ligase complex regulates endocytic trafficking by targeting EPS15 at endosomes

  1. Michaela Gschweitl
  2. Anna Ulbricht
  3. Christopher A Barnes
  4. Radoslav I Enchev
  5. Ingrid Stoffel-Studer
  6. Nathalie Meyer-Schaller
  7. Jatta Huotari
  8. Yohei Yamauchi
  9. Urs F Greber
  10. Ari Helenius
  11. Matthias Peter  Is a corresponding author
  1. Eidgenössische Technische Hochschule Zürich, Switzerland
  2. Novo Nordisk Research Center, United States
  3. University of Basel, Switzerland
  4. nspm ltd, Switzerland
  5. University of Zurich, Switzerland
Research Article
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Cite this article as: eLife 2016;5:e13841 doi: 10.7554/eLife.13841

Abstract

Cullin-3 (CUL3)-based ubiquitin ligases regulate endosome maturation and trafficking of endocytic cargo to lysosomes in mammalian cells. Here, we report that these functions depend on SPOPL, a substrate-specific CUL3 adaptor. We find that SPOPL associates with endosomes and is required for both the formation of multivesicular bodies (MVBs) and the endocytic host cell entry of influenza A virus. In SPOPL-depleted cells, endosomes are enlarged and fail to acquire intraluminal vesicles (ILVs). We identify a critical substrate ubiquitinated by CUL3-SPOPL as EPS15, an endocytic adaptor that also associates with the ESCRT-0 complex members HRS and STAM on endosomes. Indeed, EPS15 is ubiquitinated in a SPOPL-dependent manner, and accumulates with HRS in cells lacking SPOPL. Together, our data indicates that a CUL3-SPOPL E3 ubiquitin ligase complex regulates endocytic trafficking and MVB formation by ubiquitinating and degrading EPS15 at endosomes, thereby influencing influenza A virus infection as well as degradation of EGFR and other EPS15 targets.

Article and author information

Author details

  1. Michaela Gschweitl

    Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zürich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  2. Anna Ulbricht

    Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  3. Christopher A Barnes

    Novo Nordisk Research Center, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Radoslav I Enchev

    Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  5. Ingrid Stoffel-Studer

    Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  6. Nathalie Meyer-Schaller

    Department of Biomedicine, University of Basel, Basel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  7. Jatta Huotari

    nspm ltd, Meggen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  8. Yohei Yamauchi

    Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  9. Urs F Greber

    Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  10. Ari Helenius

    Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  11. Matthias Peter

    Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland
    For correspondence
    matthias.peter@bc.biol.ethz.ch
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. J Wade Harper, Harvard Medical School, United States

Publication history

  1. Received: December 16, 2015
  2. Accepted: March 23, 2016
  3. Accepted Manuscript published: March 23, 2016 (version 1)
  4. Version of Record published: April 21, 2016 (version 2)

Copyright

© 2016, Gschweitl 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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