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RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis

Breast epithelial cells cultured in three-dimensional (3D) collagen gels undergo ductal morphogenesis when the gel is compliant and they can achieve tensional homeostasis. We previously showed that this process requires down-regulation of Rho in compliant collagen gels, but the mechanism remains und...

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Autores principales: Ponik, Suzanne M., Trier, Steven M., Wozniak, Michele A., Eliceiri, Kevin W., Keely, Patricia J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3667722/
https://www.ncbi.nlm.nih.gov/pubmed/23552690
http://dx.doi.org/10.1091/mbc.E12-05-0386
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author Ponik, Suzanne M.
Trier, Steven M.
Wozniak, Michele A.
Eliceiri, Kevin W.
Keely, Patricia J.
author_facet Ponik, Suzanne M.
Trier, Steven M.
Wozniak, Michele A.
Eliceiri, Kevin W.
Keely, Patricia J.
author_sort Ponik, Suzanne M.
collection PubMed
description Breast epithelial cells cultured in three-dimensional (3D) collagen gels undergo ductal morphogenesis when the gel is compliant and they can achieve tensional homeostasis. We previously showed that this process requires down-regulation of Rho in compliant collagen gels, but the mechanism remains undefined. In this study, we find that p190RhoGAP-B, but not p190RhoGAP-A, mediates down-regulation of RhoA activity and ductal morphogenesis in T47D cells cultured in compliant 3D collagen gels. In addition, both RhoA and p190RhoGAP-B colocalize with p120-catenin at sites of cell–cell contact. The association between p190RhoGAP-B and p120-catenin is regulated by matrix compliance such that it increases in compliant vs. rigid collagen gels. Furthermore, knockdown of p120-catenin disrupts ductal morphogenesis, disregulates RhoA activity, and results in loss of p190B at cell–cell contacts. Consistent with these findings, using a RhoA-specific FRET biosensor (RhoA-FLARE.sc), we determined spatial RhoA activity to be significantly decreased at cell–cell contacts versus cell–ECM adhesions, and, of importance, spatial RhoA activity is regulated by p190B. This finding suggests that RhoA exists as an inactive pool at cell–cell contacts and is recruited to cell–ECM contacts within stiff matrices. Overall, these results demonstrate that RhoA is down-regulated at cell–cell contacts through p190RhoGAP-B, which is localized to cell–cell contacts by association with p120-catenin that is regulated by tensional homeostasis.
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spelling pubmed-36677222013-08-16 RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis Ponik, Suzanne M. Trier, Steven M. Wozniak, Michele A. Eliceiri, Kevin W. Keely, Patricia J. Mol Biol Cell Articles Breast epithelial cells cultured in three-dimensional (3D) collagen gels undergo ductal morphogenesis when the gel is compliant and they can achieve tensional homeostasis. We previously showed that this process requires down-regulation of Rho in compliant collagen gels, but the mechanism remains undefined. In this study, we find that p190RhoGAP-B, but not p190RhoGAP-A, mediates down-regulation of RhoA activity and ductal morphogenesis in T47D cells cultured in compliant 3D collagen gels. In addition, both RhoA and p190RhoGAP-B colocalize with p120-catenin at sites of cell–cell contact. The association between p190RhoGAP-B and p120-catenin is regulated by matrix compliance such that it increases in compliant vs. rigid collagen gels. Furthermore, knockdown of p120-catenin disrupts ductal morphogenesis, disregulates RhoA activity, and results in loss of p190B at cell–cell contacts. Consistent with these findings, using a RhoA-specific FRET biosensor (RhoA-FLARE.sc), we determined spatial RhoA activity to be significantly decreased at cell–cell contacts versus cell–ECM adhesions, and, of importance, spatial RhoA activity is regulated by p190B. This finding suggests that RhoA exists as an inactive pool at cell–cell contacts and is recruited to cell–ECM contacts within stiff matrices. Overall, these results demonstrate that RhoA is down-regulated at cell–cell contacts through p190RhoGAP-B, which is localized to cell–cell contacts by association with p120-catenin that is regulated by tensional homeostasis. The American Society for Cell Biology 2013-06-01 /pmc/articles/PMC3667722/ /pubmed/23552690 http://dx.doi.org/10.1091/mbc.E12-05-0386 Text en © 2013 Ponik 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 of Cell Biology.
spellingShingle Articles
Ponik, Suzanne M.
Trier, Steven M.
Wozniak, Michele A.
Eliceiri, Kevin W.
Keely, Patricia J.
RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis
title RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis
title_full RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis
title_fullStr RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis
title_full_unstemmed RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis
title_short RhoA is down-regulated at cell–cell contacts via p190RhoGAP-B in response to tensional homeostasis
title_sort rhoa is down-regulated at cell–cell contacts via p190rhogap-b in response to tensional homeostasis
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3667722/
https://www.ncbi.nlm.nih.gov/pubmed/23552690
http://dx.doi.org/10.1091/mbc.E12-05-0386
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