Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.

We developed new image analysis tools to analyse quantitatively the extracellular-matrix-dependent cell spreading process imaged by live-cell epifluorescence microscopy. Using these tools, we investigated cell spreading induced by activation of the small GTPase, Rap1. After replating and initial adh...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sarah H Ross, Emma Spanjaard, Anneke Post, Marjolein J Vliem, Hendy Kristyanto, Johannes L Bos, Johan de Rooij
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3ef73f727ea84a80bc37a1e8f771963e
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:3ef73f727ea84a80bc37a1e8f771963e
record_format	dspace
spelling	oai:doaj.org-article:3ef73f727ea84a80bc37a1e8f771963e2021-11-18T08:07:26ZRap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.1932-620310.1371/journal.pone.0050072https://doaj.org/article/3ef73f727ea84a80bc37a1e8f771963e2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23209645/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203We developed new image analysis tools to analyse quantitatively the extracellular-matrix-dependent cell spreading process imaged by live-cell epifluorescence microscopy. Using these tools, we investigated cell spreading induced by activation of the small GTPase, Rap1. After replating and initial adhesion, unstimulated cells exhibited extensive protrusion and retraction as their spread area increased, and displayed an angular shape that was remodelled over time. In contrast, activation of endogenous Rap1, via 007-mediated stimulation of Epac1, induced protrusion along the entire cell periphery, resulting in a rounder spread surface, an accelerated spreading rate and an increased spread area compared to control cells. Whereas basal, anisotropic, spreading was completely dependent on Src activity, Rap1-induced spreading was refractory to Src inhibition. Under Src inhibited conditions, the characteristic Src-induced tyrosine phosphorylations of FAK and paxillin did not occur, but Rap1 could induce the formation of actomyosin-connected adhesions, which contained vinculin at levels comparable to that found in unperturbed focal adhesions. From these results, we conclude that Rap1 can induce cell adhesion and stimulate an accelerated rate of cell spreading through mechanisms that bypass the canonical FAK-Src-Paxillin signalling cascade.Sarah H RossEmma SpanjaardAnneke PostMarjolein J VliemHendy KristyantoJohannes L BosJohan de RooijPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 11, p e50072 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sarah H Ross Emma Spanjaard Anneke Post Marjolein J Vliem Hendy Kristyanto Johannes L Bos Johan de Rooij Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.
description	We developed new image analysis tools to analyse quantitatively the extracellular-matrix-dependent cell spreading process imaged by live-cell epifluorescence microscopy. Using these tools, we investigated cell spreading induced by activation of the small GTPase, Rap1. After replating and initial adhesion, unstimulated cells exhibited extensive protrusion and retraction as their spread area increased, and displayed an angular shape that was remodelled over time. In contrast, activation of endogenous Rap1, via 007-mediated stimulation of Epac1, induced protrusion along the entire cell periphery, resulting in a rounder spread surface, an accelerated spreading rate and an increased spread area compared to control cells. Whereas basal, anisotropic, spreading was completely dependent on Src activity, Rap1-induced spreading was refractory to Src inhibition. Under Src inhibited conditions, the characteristic Src-induced tyrosine phosphorylations of FAK and paxillin did not occur, but Rap1 could induce the formation of actomyosin-connected adhesions, which contained vinculin at levels comparable to that found in unperturbed focal adhesions. From these results, we conclude that Rap1 can induce cell adhesion and stimulate an accelerated rate of cell spreading through mechanisms that bypass the canonical FAK-Src-Paxillin signalling cascade.
format	article
author	Sarah H Ross Emma Spanjaard Anneke Post Marjolein J Vliem Hendy Kristyanto Johannes L Bos Johan de Rooij
author_facet	Sarah H Ross Emma Spanjaard Anneke Post Marjolein J Vliem Hendy Kristyanto Johannes L Bos Johan de Rooij
author_sort	Sarah H Ross
title	Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.
title_short	Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.
title_full	Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.
title_fullStr	Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.
title_full_unstemmed	Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.
title_sort	rap1 can bypass the fak-src-paxillin cascade to induce cell spreading and focal adhesion formation.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/3ef73f727ea84a80bc37a1e8f771963e
work_keys_str_mv	AT sarahhross rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation AT emmaspanjaard rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation AT annekepost rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation AT marjoleinjvliem rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation AT hendykristyanto rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation AT johanneslbos rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation AT johanderooij rap1canbypassthefaksrcpaxillincascadetoinducecellspreadingandfocaladhesionformation
_version_	1718422163004850176

Rap1 can bypass the FAK-Src-Paxillin cascade to induce cell spreading and focal adhesion formation.

Ejemplares similares