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The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells

RhoGTPases are important regulators of the cell cytoskeleton, controlling cell shape, migration and proliferation. Previously we showed that ARHGAP18 in endothelial cells is important in cell junctions. Here we show, using structured illumination microscopy (SIM), ground-state depletion (GSD), and t...

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Autores principales: Lovelace, Michael D., Powter, Elizabeth E., Coleman, Paul R., Zhao, Yang, Parker, Amelia, Chang, Garry H., Lay, Angelina J., Hunter, Julie, McGrath, Aaron P., Jormakka, Mika, Bertolino, Patrick, McCaughan, Geoffrey, Kavallaris, Maria, Vadas, Mathew A., Gamble, Jennifer R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5391183/
https://www.ncbi.nlm.nih.gov/pubmed/28251925
http://dx.doi.org/10.1091/mbc.E16-05-0285
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author Lovelace, Michael D.
Powter, Elizabeth E.
Coleman, Paul R.
Zhao, Yang
Parker, Amelia
Chang, Garry H.
Lay, Angelina J.
Hunter, Julie
McGrath, Aaron P.
Jormakka, Mika
Bertolino, Patrick
McCaughan, Geoffrey
Kavallaris, Maria
Vadas, Mathew A.
Gamble, Jennifer R.
author_facet Lovelace, Michael D.
Powter, Elizabeth E.
Coleman, Paul R.
Zhao, Yang
Parker, Amelia
Chang, Garry H.
Lay, Angelina J.
Hunter, Julie
McGrath, Aaron P.
Jormakka, Mika
Bertolino, Patrick
McCaughan, Geoffrey
Kavallaris, Maria
Vadas, Mathew A.
Gamble, Jennifer R.
author_sort Lovelace, Michael D.
collection PubMed
description RhoGTPases are important regulators of the cell cytoskeleton, controlling cell shape, migration and proliferation. Previously we showed that ARHGAP18 in endothelial cells is important in cell junctions. Here we show, using structured illumination microscopy (SIM), ground-state depletion (GSD), and total internal reflection fluorescence microscopy (TIRF) that a proportion of ARHGAP18 localizes to microtubules in endothelial cells, as well as in nonendothelial cells, an association confirmed biochemically. In endothelial cells, some ARHGAP18 puncta also colocalized to Weibel–Palade bodies on the microtubules. Depletion of ARHGAP18 by small interfering RNA or analysis of endothelial cells isolated from ARHGAP18-knockout mice showed microtubule destabilization, as evidenced by altered morphology and decreased acetylated α-tubulin and glu-tubulin. The destabilization was rescued by inhibition of ROCK and histone deacetylase 6 but not by a GAP-mutant form of ARHGAP18. Depletion of ARHGAP18 resulted in a failure to secrete endothelin-1 and a reduction in neutrophil transmigration, both known to be microtubule dependent. Thrombin, a critical regulator of the Rho-mediated barrier function of endothelial cells through microtubule destabilization, enhanced the plasma membrane–bound fraction of ARHGAP18. Thus, in endothelial cells, ARHGAP18 may act as a significant regulator of vascular homeostasis.
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spelling pubmed-53911832017-06-30 The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells Lovelace, Michael D. Powter, Elizabeth E. Coleman, Paul R. Zhao, Yang Parker, Amelia Chang, Garry H. Lay, Angelina J. Hunter, Julie McGrath, Aaron P. Jormakka, Mika Bertolino, Patrick McCaughan, Geoffrey Kavallaris, Maria Vadas, Mathew A. Gamble, Jennifer R. Mol Biol Cell Articles RhoGTPases are important regulators of the cell cytoskeleton, controlling cell shape, migration and proliferation. Previously we showed that ARHGAP18 in endothelial cells is important in cell junctions. Here we show, using structured illumination microscopy (SIM), ground-state depletion (GSD), and total internal reflection fluorescence microscopy (TIRF) that a proportion of ARHGAP18 localizes to microtubules in endothelial cells, as well as in nonendothelial cells, an association confirmed biochemically. In endothelial cells, some ARHGAP18 puncta also colocalized to Weibel–Palade bodies on the microtubules. Depletion of ARHGAP18 by small interfering RNA or analysis of endothelial cells isolated from ARHGAP18-knockout mice showed microtubule destabilization, as evidenced by altered morphology and decreased acetylated α-tubulin and glu-tubulin. The destabilization was rescued by inhibition of ROCK and histone deacetylase 6 but not by a GAP-mutant form of ARHGAP18. Depletion of ARHGAP18 resulted in a failure to secrete endothelin-1 and a reduction in neutrophil transmigration, both known to be microtubule dependent. Thrombin, a critical regulator of the Rho-mediated barrier function of endothelial cells through microtubule destabilization, enhanced the plasma membrane–bound fraction of ARHGAP18. Thus, in endothelial cells, ARHGAP18 may act as a significant regulator of vascular homeostasis. The American Society for Cell Biology 2017-04-15 /pmc/articles/PMC5391183/ /pubmed/28251925 http://dx.doi.org/10.1091/mbc.E16-05-0285 Text en © 2017 Lovelace et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Lovelace, Michael D.
Powter, Elizabeth E.
Coleman, Paul R.
Zhao, Yang
Parker, Amelia
Chang, Garry H.
Lay, Angelina J.
Hunter, Julie
McGrath, Aaron P.
Jormakka, Mika
Bertolino, Patrick
McCaughan, Geoffrey
Kavallaris, Maria
Vadas, Mathew A.
Gamble, Jennifer R.
The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells
title The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells
title_full The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells
title_fullStr The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells
title_full_unstemmed The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells
title_short The RhoGAP protein ARHGAP18/SENEX localizes to microtubules and regulates their stability in endothelial cells
title_sort rhogap protein arhgap18/senex localizes to microtubules and regulates their stability in endothelial cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5391183/
https://www.ncbi.nlm.nih.gov/pubmed/28251925
http://dx.doi.org/10.1091/mbc.E16-05-0285
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