1. Biophysics and Structural Biology
  2. Cell Biology
Download icon

Visualizing the functional architecture of the endocytic machinery

  1. Andrea Picco
  2. Markus Mund
  3. Jonas Ries
  4. François Nédélec
  5. Marko Kaksonen Is a corresponding author
  1. European Molecular Biology Laboratory, Germany
Research Article
Cited
13
Views
4,703
Comments
0
Cite as: eLife 2015;4:e04535 doi: 10.7554/eLife.04535

Abstract

Clathrin-mediated endocytosis is an essential process that forms vesicles from the plasma membrane. Although most of the protein components of the endocytic protein machinery have been thoroughly characterized, their organization at the endocytic site is poorly understood. We developed a fluorescence microscopy method to track the average positions of yeast endocytic proteins in relation to each other with a time precision below 1 s and with a spatial precision of ~10 nm. With these data, integrated with shapes of endocytic membrane intermediates and with superresolution imaging, we could visualize the dynamic architecture of the endocytic machinery. We showed how different coat proteins are distributed within the coat structure and how the assembly dynamics of N-BAR proteins relate to membrane shape changes. Moreover, we found that the region of actin polymerization is located at the base of the endocytic invagination, with the growing ends of filaments pointing toward the plasma membrane.

Article and author information

Author details

  1. Andrea Picco

    1. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Markus Mund

    1. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Jonas Ries

    1. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. François Nédélec

    1. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Marko Kaksonen

    1. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    For correspondence
    1. kaksonen@embl.de
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Suzanne R Pfeffer, Reviewing Editor, Stanford University, United States

Publication history

  1. Received: August 28, 2014
  2. Accepted: February 10, 2015
  3. Accepted Manuscript published: February 12, 2015 (version 1)
  4. Version of Record published: March 12, 2015 (version 2)

Copyright

© 2015, Picco 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

  • 4,703
    Page views
  • 921
    Downloads
  • 13
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Comments

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. Biophysics and Structural Biology
    2. Microbiology and Infectious Disease
    Mauricio Comas-Garcia et al.
    Research Article
    1. Developmental Biology and Stem Cells
    Cyrille Ramond et al.
    Research Article