Epsin deficiency impairs endocytosis by stalling the actin-dependent invagination of endocytic clathrin-coated pits

  1. Mirko Messa
  2. Ruben Fernandez-Busnadiego
  3. Elizabeth Wen Sun
  4. Hong Chen
  5. Heather Czapla
  6. Kristie Wrasman
  7. Yumei Wu
  8. Genevieve Ko
  9. Theodora Ross
  10. Beverly Wendland
  11. Pietro De Camilli  Is a corresponding author
  1. Howard Hughes Medical Institute, Yale University School of Medicine, United States
  2. Max-Planck-Institut für Biochemie, Germany
  3. Oklahoma Medical Research Foundation, United States
  4. Johns Hopkins University, United States
  5. UT Southwestern Medical Center, United States

Abstract

Epsin is an evolutionarily conserved endocytic clathrin adaptor whose most critical function(s) in clathrin coat dynamics remain(s) elusive. To elucidate such function(s), we generated embryonic fibroblasts from conditional epsin triple KO mice. Triple KO cells displayed a dramatic cell division defect. Additionally, a robust impairment in clathrin-mediated endocytosis was observed, with an accumulation of early and U-shaped pits. This defect correlated with a perturbation of the coupling between the clathrin coat and the actin cytoskeleton, which we confirmed in a cell-free assay of endocytosis. Our results indicate that a key evolutionary conserved function of epsin, in addition to other roles that include as we show here a low affinity interaction with SNAREs, is to help generate the force that leads to invagination and then fission of clathrin-coated pits.

Article and author information

Author details

  1. Mirko Messa

    Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Ruben Fernandez-Busnadiego

    Max-Planck-Institut für Biochemie, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Elizabeth Wen Sun

    Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Hong Chen

    Oklahoma Medical Research Foundation, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Heather Czapla

    Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Kristie Wrasman

    Johns Hopkins University, Baltimore, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Yumei Wu

    Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Genevieve Ko

    Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Theodora Ross

    UT Southwestern Medical Center, Dallas, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Beverly Wendland

    Johns Hopkins University, Baltimore, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Pietro De Camilli

    Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States
    For correspondence
    pietro.decamilli@yale.edu
    Competing interests
    The authors declare that no competing interests exist.

Ethics

Animal experimentation: The institutional animal care and use committee (IACUC) of the Yale University and the approved animal protocol is 2012-07422. The institutional guidelines for the care and use of laboratory animals were followed

Reviewing Editor

  1. Suzanne R Pfeffer, Stanford University, United States

Publication history

  1. Received: May 7, 2014
  2. Accepted: August 12, 2014
  3. Accepted Manuscript published: August 13, 2014 (version 1)
  4. Version of Record published: September 12, 2014 (version 2)

Copyright

© 2014, Messa 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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  1. Mirko Messa
  2. Ruben Fernandez-Busnadiego
  3. Elizabeth Wen Sun
  4. Hong Chen
  5. Heather Czapla
  6. Kristie Wrasman
  7. Yumei Wu
  8. Genevieve Ko
  9. Theodora Ross
  10. Beverly Wendland
  11. Pietro De Camilli
(2014)
Epsin deficiency impairs endocytosis by stalling the actin-dependent invagination of endocytic clathrin-coated pits
eLife 3:e03311.
https://doi.org/10.7554/eLife.03311

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