Cargando…

Visualizing the functional architecture of the endocytic machinery

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 fluorescenc...

Descripción completa

Detalles Bibliográficos
Autores principales: Picco, Andrea, Mund, Markus, Ries, Jonas, Nédélec, François, Kaksonen, Marko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357291/
https://www.ncbi.nlm.nih.gov/pubmed/25675087
http://dx.doi.org/10.7554/eLife.04535
_version_ 1782361129400401920
author Picco, Andrea
Mund, Markus
Ries, Jonas
Nédélec, François
Kaksonen, Marko
author_facet Picco, Andrea
Mund, Markus
Ries, Jonas
Nédélec, François
Kaksonen, Marko
author_sort Picco, Andrea
collection PubMed
description 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. DOI: http://dx.doi.org/10.7554/eLife.04535.001
format Online
Article
Text
id pubmed-4357291
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher eLife Sciences Publications, Ltd
record_format MEDLINE/PubMed
spelling pubmed-43572912015-03-16 Visualizing the functional architecture of the endocytic machinery Picco, Andrea Mund, Markus Ries, Jonas Nédélec, François Kaksonen, Marko eLife Biophysics and Structural Biology 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. DOI: http://dx.doi.org/10.7554/eLife.04535.001 eLife Sciences Publications, Ltd 2015-02-12 /pmc/articles/PMC4357291/ /pubmed/25675087 http://dx.doi.org/10.7554/eLife.04535 Text en © 2015, Picco et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biophysics and Structural Biology
Picco, Andrea
Mund, Markus
Ries, Jonas
Nédélec, François
Kaksonen, Marko
Visualizing the functional architecture of the endocytic machinery
title Visualizing the functional architecture of the endocytic machinery
title_full Visualizing the functional architecture of the endocytic machinery
title_fullStr Visualizing the functional architecture of the endocytic machinery
title_full_unstemmed Visualizing the functional architecture of the endocytic machinery
title_short Visualizing the functional architecture of the endocytic machinery
title_sort visualizing the functional architecture of the endocytic machinery
topic Biophysics and Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357291/
https://www.ncbi.nlm.nih.gov/pubmed/25675087
http://dx.doi.org/10.7554/eLife.04535
work_keys_str_mv AT piccoandrea visualizingthefunctionalarchitectureoftheendocyticmachinery
AT mundmarkus visualizingthefunctionalarchitectureoftheendocyticmachinery
AT riesjonas visualizingthefunctionalarchitectureoftheendocyticmachinery
AT nedelecfrancois visualizingthefunctionalarchitectureoftheendocyticmachinery
AT kaksonenmarko visualizingthefunctionalarchitectureoftheendocyticmachinery