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EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress

Caveolae introduce flask-shaped convolutions into the plasma membrane and help to protect the plasma membrane from damage under stretch forces. The protein components that form the bulb of caveolae are increasingly well characterized, but less is known about the contribution of proteins that localiz...

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Autores principales: Yeow, Ivana, Howard, Gillian, Chadwick, Jessica, Mendoza-Topaz, Carolina, Hansen, Carsten G., Nichols, Benjamin J., Shvets, Elena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5640515/
https://www.ncbi.nlm.nih.gov/pubmed/28943089
http://dx.doi.org/10.1016/j.cub.2017.07.047
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author Yeow, Ivana
Howard, Gillian
Chadwick, Jessica
Mendoza-Topaz, Carolina
Hansen, Carsten G.
Nichols, Benjamin J.
Shvets, Elena
author_facet Yeow, Ivana
Howard, Gillian
Chadwick, Jessica
Mendoza-Topaz, Carolina
Hansen, Carsten G.
Nichols, Benjamin J.
Shvets, Elena
author_sort Yeow, Ivana
collection PubMed
description Caveolae introduce flask-shaped convolutions into the plasma membrane and help to protect the plasma membrane from damage under stretch forces. The protein components that form the bulb of caveolae are increasingly well characterized, but less is known about the contribution of proteins that localize to the constricted neck. Here we make extensive use of multiple CRISPR/Cas9-generated gene knockout and knockin cell lines to investigate the role of Eps15 Homology Domain (EHD) proteins at the neck of caveolae. We show that EHD1, EHD2, and EHD4 are recruited to caveolae. Recruitment of the other EHDs increases markedly when EHD2, which has been previously detected at caveolae, is absent. Construction of knockout cell lines lacking EHDs 1, 2, and 4 confirms this apparent functional redundancy. Two striking sets of phenotypes are observed in EHD1,2,4 knockout cells: (1) the characteristic clustering of caveolae into higher-order assemblies is absent; and (2) when the EHD1,2,4 knockout cells are subjected to prolonged cycles of stretch forces, caveolae are destabilized and the plasma membrane is prone to rupture. Our data identify the first molecular components that act to cluster caveolae into a membrane ultrastructure with the potential to extend stretch-buffering capacity and support a revised model for the function of EHDs at the caveolar neck.
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spelling pubmed-56405152017-10-20 EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress Yeow, Ivana Howard, Gillian Chadwick, Jessica Mendoza-Topaz, Carolina Hansen, Carsten G. Nichols, Benjamin J. Shvets, Elena Curr Biol Article Caveolae introduce flask-shaped convolutions into the plasma membrane and help to protect the plasma membrane from damage under stretch forces. The protein components that form the bulb of caveolae are increasingly well characterized, but less is known about the contribution of proteins that localize to the constricted neck. Here we make extensive use of multiple CRISPR/Cas9-generated gene knockout and knockin cell lines to investigate the role of Eps15 Homology Domain (EHD) proteins at the neck of caveolae. We show that EHD1, EHD2, and EHD4 are recruited to caveolae. Recruitment of the other EHDs increases markedly when EHD2, which has been previously detected at caveolae, is absent. Construction of knockout cell lines lacking EHDs 1, 2, and 4 confirms this apparent functional redundancy. Two striking sets of phenotypes are observed in EHD1,2,4 knockout cells: (1) the characteristic clustering of caveolae into higher-order assemblies is absent; and (2) when the EHD1,2,4 knockout cells are subjected to prolonged cycles of stretch forces, caveolae are destabilized and the plasma membrane is prone to rupture. Our data identify the first molecular components that act to cluster caveolae into a membrane ultrastructure with the potential to extend stretch-buffering capacity and support a revised model for the function of EHDs at the caveolar neck. Cell Press 2017-10-09 /pmc/articles/PMC5640515/ /pubmed/28943089 http://dx.doi.org/10.1016/j.cub.2017.07.047 Text en © 2017 MRC Laboratory of Molecular Biology http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yeow, Ivana
Howard, Gillian
Chadwick, Jessica
Mendoza-Topaz, Carolina
Hansen, Carsten G.
Nichols, Benjamin J.
Shvets, Elena
EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress
title EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress
title_full EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress
title_fullStr EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress
title_full_unstemmed EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress
title_short EHD Proteins Cooperate to Generate Caveolar Clusters and to Maintain Caveolae during Repeated Mechanical Stress
title_sort ehd proteins cooperate to generate caveolar clusters and to maintain caveolae during repeated mechanical stress
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5640515/
https://www.ncbi.nlm.nih.gov/pubmed/28943089
http://dx.doi.org/10.1016/j.cub.2017.07.047
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