Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains

Abstract Cell motility and migration requires the reorganization of the cortical cytoskeleton at the leading edge of cells and extracellular Ca2+ entry is essential for this reorganization. However the molecular nature of the regulators of this pathway is unknown. This work contributes to understand...

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Autores principales:	Aida M. Lopez-Guerrero, Patricia Tomas-Martin, Carlos Pascual-Caro, Thomas Macartney, Alejandro Rojas-Fernandez, Graeme Ball, Dario R. Alessi, Eulalia Pozo-Guisado, Francisco Javier Martin-Romero
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Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/def4895300394d3fa46a7edb9c8abe1b
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spelling	oai:doaj.org-article:def4895300394d3fa46a7edb9c8abe1b2021-12-02T16:06:56ZRegulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains10.1038/s41598-017-00331-42045-2322https://doaj.org/article/def4895300394d3fa46a7edb9c8abe1b2017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00331-4https://doaj.org/toc/2045-2322Abstract Cell motility and migration requires the reorganization of the cortical cytoskeleton at the leading edge of cells and extracellular Ca2+ entry is essential for this reorganization. However the molecular nature of the regulators of this pathway is unknown. This work contributes to understanding the role of STIM1 and ORAI1 in the promotion of membrane ruffling by showing that phospho-STIM1 localizes at the leading edge of cells, and that both phospho-STIM1 and ORAI1 co-localize with cortactin (CTTN), a regulator of the cytoskeleton at membrane ruffling areas. STIM1-KO and ORAI1-KO cell lines were generated by CRISPR/Cas9 genome editing in U2OS cells. In both cases, KO cells presented a notable reduction of store-operated Ca2+ entry (SOCE) that was rescued by expression of STIM1-mCherry and ORAI1-mCherry. These results demonstrated that SOCE regulates membrane ruffling at the leading edge of cells. Moreover, endogenous ORAI1 and overexpressed ORAI1-GFP co-immunoprecipitated with endogenous CTTN. This latter result, in addition to the KO cells’ phenotype, the preservation of ORAI1-CTTN co-localization during ruffling, and the inhibition of membrane ruffling by the Ca2+-channel inhibitor SKF96365, further supports a functional link between SOCE and membrane ruffling.Aida M. Lopez-GuerreroPatricia Tomas-MartinCarlos Pascual-CaroThomas MacartneyAlejandro Rojas-FernandezGraeme BallDario R. AlessiEulalia Pozo-GuisadoFrancisco Javier Martin-RomeroNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-15 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Aida M. Lopez-Guerrero Patricia Tomas-Martin Carlos Pascual-Caro Thomas Macartney Alejandro Rojas-Fernandez Graeme Ball Dario R. Alessi Eulalia Pozo-Guisado Francisco Javier Martin-Romero Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
description	Abstract Cell motility and migration requires the reorganization of the cortical cytoskeleton at the leading edge of cells and extracellular Ca2+ entry is essential for this reorganization. However the molecular nature of the regulators of this pathway is unknown. This work contributes to understanding the role of STIM1 and ORAI1 in the promotion of membrane ruffling by showing that phospho-STIM1 localizes at the leading edge of cells, and that both phospho-STIM1 and ORAI1 co-localize with cortactin (CTTN), a regulator of the cytoskeleton at membrane ruffling areas. STIM1-KO and ORAI1-KO cell lines were generated by CRISPR/Cas9 genome editing in U2OS cells. In both cases, KO cells presented a notable reduction of store-operated Ca2+ entry (SOCE) that was rescued by expression of STIM1-mCherry and ORAI1-mCherry. These results demonstrated that SOCE regulates membrane ruffling at the leading edge of cells. Moreover, endogenous ORAI1 and overexpressed ORAI1-GFP co-immunoprecipitated with endogenous CTTN. This latter result, in addition to the KO cells’ phenotype, the preservation of ORAI1-CTTN co-localization during ruffling, and the inhibition of membrane ruffling by the Ca2+-channel inhibitor SKF96365, further supports a functional link between SOCE and membrane ruffling.
format	article
author	Aida M. Lopez-Guerrero Patricia Tomas-Martin Carlos Pascual-Caro Thomas Macartney Alejandro Rojas-Fernandez Graeme Ball Dario R. Alessi Eulalia Pozo-Guisado Francisco Javier Martin-Romero
author_facet	Aida M. Lopez-Guerrero Patricia Tomas-Martin Carlos Pascual-Caro Thomas Macartney Alejandro Rojas-Fernandez Graeme Ball Dario R. Alessi Eulalia Pozo-Guisado Francisco Javier Martin-Romero
author_sort	Aida M. Lopez-Guerrero
title	Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
title_short	Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
title_full	Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
title_fullStr	Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
title_full_unstemmed	Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
title_sort	regulation of membrane ruffling by polarized stim1 and orai1 in cortactin-rich domains
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/def4895300394d3fa46a7edb9c8abe1b
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Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains

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