Single-molecule turnover dynamics of actin and membrane coat proteins in clathrin-mediated endocytosis

  1. Michael M Lacy
  2. David Baddeley
  3. Julien Berro  Is a corresponding author
  1. Yale University, United States

Abstract

Actin dynamics generate forces to deform the membrane and overcome the cell's high turgor pressure during clathrin-mediated endocytosis (CME) in yeast, but precise molecular details are still unresolved. Our previous models predicted that actin filaments of the endocytic meshwork continually polymerize and disassemble, turning over multiple times during an endocytic event, similar to other actin systems. We applied single-molecule speckle tracking in live fission yeast to directly measure molecular turnover within CME sites for the first time. In contrast with the overall ~20-sec lifetimes of actin and actin-associated proteins in endocytic patches, we detected single-molecule residence times around 1 to 2 sec, and similarly high turnover rates of membrane-associated proteins in CME. Furthermore, we find heterogeneous behaviors in many proteins' motions. These results indicate that endocytic proteins turn over up to five times during the formation of an endocytic vesicle, and suggest revising quantitative models of force production.

Data availability

All data generated or analysed during this study are included in the manuscript and the supporting file "SuppFile4sets.mat" (Matlab data file).

Article and author information

Author details

  1. Michael M Lacy

    Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0498-2817
  2. David Baddeley

    Nanobiology Institute, Yale University, West Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Julien Berro

    Department of Molecular Biophysics and Biochemistry, Department of Cell Biology, Yale University, New Haven, United States
    For correspondence
    julien.berro@yale.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9560-8646

Funding

National Institute of General Medical Sciences (R01GM115636)

  • Michael M Lacy
  • Julien Berro

National Institute of General Medical Sciences (T32GM008283)

  • Michael M Lacy

Yale Program in Physics Engineering and Biology

  • Michael M Lacy
  • David Baddeley
  • Julien Berro

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

Reviewing Editor

  1. Pekka Lappalainen, University of Helsinki, Finland

Version history

  1. Received: October 3, 2019
  2. Accepted: December 18, 2019
  3. Accepted Manuscript published: December 19, 2019 (version 1)
  4. Version of Record published: January 23, 2020 (version 2)

Copyright

© 2019, Lacy 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. Michael M Lacy
  2. David Baddeley
  3. Julien Berro
(2019)
Single-molecule turnover dynamics of actin and membrane coat proteins in clathrin-mediated endocytosis
eLife 8:e52355.
https://doi.org/10.7554/eLife.52355

Share this article

https://doi.org/10.7554/eLife.52355

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