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Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization

The small GTPase RhoA is involved in cell morphology and migration. RhoA activity is tightly regulated in time and space and depends on guanine exchange factors (GEFs). However, the kinetics and subcellular localization of GEF activity towards RhoA are poorly defined. To study the mechanism underlyi...

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Autores principales: van Unen, Jakobus, Reinhard, Nathalie R., Yin, Taofei, Wu, Yi I., Postma, Marten, Gadella, Theodorus W.J., Goedhart, Joachim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4592971/
https://www.ncbi.nlm.nih.gov/pubmed/26435194
http://dx.doi.org/10.1038/srep14693
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author van Unen, Jakobus
Reinhard, Nathalie R.
Yin, Taofei
Wu, Yi I.
Postma, Marten
Gadella, Theodorus W.J.
Goedhart, Joachim
author_facet van Unen, Jakobus
Reinhard, Nathalie R.
Yin, Taofei
Wu, Yi I.
Postma, Marten
Gadella, Theodorus W.J.
Goedhart, Joachim
author_sort van Unen, Jakobus
collection PubMed
description The small GTPase RhoA is involved in cell morphology and migration. RhoA activity is tightly regulated in time and space and depends on guanine exchange factors (GEFs). However, the kinetics and subcellular localization of GEF activity towards RhoA are poorly defined. To study the mechanism underlying the spatiotemporal control of RhoA activity by GEFs, we performed single cell imaging with an improved FRET sensor reporting on the nucleotide loading state of RhoA. By employing the FRET sensor we show that a plasma membrane located RhoGEF, p63RhoGEF, can rapidly activate RhoA through endogenous GPCRs and that localized RhoA activity at the cell periphery correlates with actin polymerization. Moreover, synthetic recruitment of the catalytic domain derived from p63RhoGEF to the plasma membrane, but not to the Golgi apparatus, is sufficient to activate RhoA. The synthetic system enables local activation of endogenous RhoA and effectively induces actin polymerization and changes in cellular morphology. Together, our data demonstrate that GEF activity at the plasma membrane is sufficient for actin polymerization via local RhoA signaling.
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spelling pubmed-45929712015-10-19 Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization van Unen, Jakobus Reinhard, Nathalie R. Yin, Taofei Wu, Yi I. Postma, Marten Gadella, Theodorus W.J. Goedhart, Joachim Sci Rep Article The small GTPase RhoA is involved in cell morphology and migration. RhoA activity is tightly regulated in time and space and depends on guanine exchange factors (GEFs). However, the kinetics and subcellular localization of GEF activity towards RhoA are poorly defined. To study the mechanism underlying the spatiotemporal control of RhoA activity by GEFs, we performed single cell imaging with an improved FRET sensor reporting on the nucleotide loading state of RhoA. By employing the FRET sensor we show that a plasma membrane located RhoGEF, p63RhoGEF, can rapidly activate RhoA through endogenous GPCRs and that localized RhoA activity at the cell periphery correlates with actin polymerization. Moreover, synthetic recruitment of the catalytic domain derived from p63RhoGEF to the plasma membrane, but not to the Golgi apparatus, is sufficient to activate RhoA. The synthetic system enables local activation of endogenous RhoA and effectively induces actin polymerization and changes in cellular morphology. Together, our data demonstrate that GEF activity at the plasma membrane is sufficient for actin polymerization via local RhoA signaling. Nature Publishing Group 2015-10-05 /pmc/articles/PMC4592971/ /pubmed/26435194 http://dx.doi.org/10.1038/srep14693 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
van Unen, Jakobus
Reinhard, Nathalie R.
Yin, Taofei
Wu, Yi I.
Postma, Marten
Gadella, Theodorus W.J.
Goedhart, Joachim
Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization
title Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization
title_full Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization
title_fullStr Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization
title_full_unstemmed Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization
title_short Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization
title_sort plasma membrane restricted rhogef activity is sufficient for rhoa-mediated actin polymerization
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4592971/
https://www.ncbi.nlm.nih.gov/pubmed/26435194
http://dx.doi.org/10.1038/srep14693
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