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Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly
Here we demonstrate that Arp2/3 regulates a transition between mesenchymal and amoeboid protrusions in MCF10A epithelial cells. Using genetic and pharmacological means, we first show Arp2/3 inhibition impairs directed cell migration. Arp2/3 inhibition results in a dramatically impaired cell adhesion...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4072770/ https://www.ncbi.nlm.nih.gov/pubmed/24967897 http://dx.doi.org/10.1371/journal.pone.0100943 |
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author | Beckham, Yvonne Vasquez, Robert J. Stricker, Jonathan Sayegh, Kareem Campillo, Clement Gardel, Margaret L. |
author_facet | Beckham, Yvonne Vasquez, Robert J. Stricker, Jonathan Sayegh, Kareem Campillo, Clement Gardel, Margaret L. |
author_sort | Beckham, Yvonne |
collection | PubMed |
description | Here we demonstrate that Arp2/3 regulates a transition between mesenchymal and amoeboid protrusions in MCF10A epithelial cells. Using genetic and pharmacological means, we first show Arp2/3 inhibition impairs directed cell migration. Arp2/3 inhibition results in a dramatically impaired cell adhesion, causing deficient cell attachment and spreading to ECM as well as an 8-fold decrease in nascent adhesion assembly at the leading edge. While Arp2/3 does not play a significant role in myosin-dependent adhesion growth, mature focal adhesions undergo large scale movements against the ECM suggesting reduced coupling to the ECM. Cell edge protrusions occur at similar rates when Arp2/3 is inhibited but their morphology is dramatically altered. Persistent lamellipodia are abrogated and we observe a markedly increased incidence of blebbing and unstable pseuodopods. Micropipette-aspiration assays indicate that Arp2/3-inhibited cells have a weak coupling between the cell cortex and the plasma membrane, and suggest a potential mechanism for increased pseudopod and bleb formation. Pseudopods are not sensitive to reduced in formin or myosin II activity. Collectively, these results indicate that Arp2/3 is not necessary for rapid protrusion of the cell edge but plays a crucial role in assembling focal adhesions required for its stabilization. |
format | Online Article Text |
id | pubmed-4072770 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-40727702014-07-02 Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly Beckham, Yvonne Vasquez, Robert J. Stricker, Jonathan Sayegh, Kareem Campillo, Clement Gardel, Margaret L. PLoS One Research Article Here we demonstrate that Arp2/3 regulates a transition between mesenchymal and amoeboid protrusions in MCF10A epithelial cells. Using genetic and pharmacological means, we first show Arp2/3 inhibition impairs directed cell migration. Arp2/3 inhibition results in a dramatically impaired cell adhesion, causing deficient cell attachment and spreading to ECM as well as an 8-fold decrease in nascent adhesion assembly at the leading edge. While Arp2/3 does not play a significant role in myosin-dependent adhesion growth, mature focal adhesions undergo large scale movements against the ECM suggesting reduced coupling to the ECM. Cell edge protrusions occur at similar rates when Arp2/3 is inhibited but their morphology is dramatically altered. Persistent lamellipodia are abrogated and we observe a markedly increased incidence of blebbing and unstable pseuodopods. Micropipette-aspiration assays indicate that Arp2/3-inhibited cells have a weak coupling between the cell cortex and the plasma membrane, and suggest a potential mechanism for increased pseudopod and bleb formation. Pseudopods are not sensitive to reduced in formin or myosin II activity. Collectively, these results indicate that Arp2/3 is not necessary for rapid protrusion of the cell edge but plays a crucial role in assembling focal adhesions required for its stabilization. Public Library of Science 2014-06-26 /pmc/articles/PMC4072770/ /pubmed/24967897 http://dx.doi.org/10.1371/journal.pone.0100943 Text en © 2014 Beckham et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Beckham, Yvonne Vasquez, Robert J. Stricker, Jonathan Sayegh, Kareem Campillo, Clement Gardel, Margaret L. Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly |
title | Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly |
title_full | Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly |
title_fullStr | Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly |
title_full_unstemmed | Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly |
title_short | Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly |
title_sort | arp2/3 inhibition induces amoeboid-like protrusions in mcf10a epithelial cells by reduced cytoskeletal-membrane coupling and focal adhesion assembly |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4072770/ https://www.ncbi.nlm.nih.gov/pubmed/24967897 http://dx.doi.org/10.1371/journal.pone.0100943 |
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