1. Structural Biology and Molecular Biophysics
  2. Cell Biology

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
https://doi.org/10.7554/eLife.04535
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  1. Andrea Picco
  2. Markus Mund
  3. Jonas Ries
  4. François Nédélec
  5. Marko Kaksonen
(2015)
Visualizing the functional architecture of the endocytic machinery
eLife 4:e04535.
https://doi.org/10.7554/eLife.04535
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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.

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Author details

  1. Andrea Picco

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

  2. Markus Mund

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

  3. Jonas Ries

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

  4. François Nédélec

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

  5. Marko Kaksonen

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

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.

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  1. Andrea Picco
  2. Markus Mund
  3. Jonas Ries
  4. François Nédélec
  5. Marko Kaksonen
(2015)
Visualizing the functional architecture of the endocytic machinery
eLife 4:e04535.
https://doi.org/10.7554/eLife.04535

Share this article

https://doi.org/10.7554/eLife.04535

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Further reading

    1. Cell Biology

    Clathrin modulates vesicle scission, but not invagination shape, in yeast endocytosis

    Wanda Kukulski, Andrea Picco ... Marko Kaksonen
    Research Advance Updated

    In a previous paper (Picco et al., 2015), the dynamic architecture of the protein machinery during clathrin-mediated endocytosis was visualized using a new live imaging and particle tracking method. Here, by combining this approach with correlative light and electron microscopy, we address the role of clathrin in this process. During endocytosis, clathrin forms a cage-like coat around the membrane and associated protein components. There is growing evidence that clathrin does not determine the membrane morphology of the invagination but rather modulates the progression of endocytosis. We investigate how the deletion of clathrin heavy chain impairs the dynamics and the morphology of the endocytic membrane in budding yeast. Our results show that clathrin is not required for elongating or shaping the endocytic membrane invagination. Instead, we find that clathrin contributes to the regularity of vesicle scission and thereby to controlling vesicle size.