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Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin
The dynamic functional linkage of cadherins with the underlying actin cytoskeleton is tightly regulated to achieve proper cell–cell adhesion. p120-catenin (p120) regulates both cadherin stability and actin dynamics, but the relationship between these two functions remains unclear. Using a novel prot...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3248908/ https://www.ncbi.nlm.nih.gov/pubmed/22031287 http://dx.doi.org/10.1091/mbc.E11-06-0497 |
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author | Smith, Andrew L. Dohn, Michael R. Brown, Meredith V. Reynolds, Albert B. |
author_facet | Smith, Andrew L. Dohn, Michael R. Brown, Meredith V. Reynolds, Albert B. |
author_sort | Smith, Andrew L. |
collection | PubMed |
description | The dynamic functional linkage of cadherins with the underlying actin cytoskeleton is tightly regulated to achieve proper cell–cell adhesion. p120-catenin (p120) regulates both cadherin stability and actin dynamics, but the relationship between these two functions remains unclear. Using a novel proteomic approach called reversible cross-link immunoprecipitation, or ReCLIP, we previously identified a physical interaction between p120 and Rho-associated protein kinase 1 (ROCK1), a major effector of RhoA. In this paper, we show that a discrete fraction of cellular ROCK1 coimmunoprecipitates with p120 and precisely colocalizes to adherens junctions (AJs). Manipulation of AJs using a calcium-switch assay and cadherin-blocking antibodies indicates direct recruitment of ROCK1 to newly forming junctions. Importantly, we find that p120 links ROCK1 to the cadherin complex, as ROCK1 coimmunoprecipitates with wild-type but not p120-uncoupled E-cadherin. Moreover, depletion of ROCK1 using short-hairpin RNA results in dramatic mislocalization of the cadherin complex and junctional actin. These data are consistent with a model in which p120 dynamically regulates Rho-GTPase activity at the cadherin complex through transient interaction with several of its up- and downstream effectors, including ROCK1. |
format | Online Article Text |
id | pubmed-3248908 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-32489082012-03-16 Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin Smith, Andrew L. Dohn, Michael R. Brown, Meredith V. Reynolds, Albert B. Mol Biol Cell Articles The dynamic functional linkage of cadherins with the underlying actin cytoskeleton is tightly regulated to achieve proper cell–cell adhesion. p120-catenin (p120) regulates both cadherin stability and actin dynamics, but the relationship between these two functions remains unclear. Using a novel proteomic approach called reversible cross-link immunoprecipitation, or ReCLIP, we previously identified a physical interaction between p120 and Rho-associated protein kinase 1 (ROCK1), a major effector of RhoA. In this paper, we show that a discrete fraction of cellular ROCK1 coimmunoprecipitates with p120 and precisely colocalizes to adherens junctions (AJs). Manipulation of AJs using a calcium-switch assay and cadherin-blocking antibodies indicates direct recruitment of ROCK1 to newly forming junctions. Importantly, we find that p120 links ROCK1 to the cadherin complex, as ROCK1 coimmunoprecipitates with wild-type but not p120-uncoupled E-cadherin. Moreover, depletion of ROCK1 using short-hairpin RNA results in dramatic mislocalization of the cadherin complex and junctional actin. These data are consistent with a model in which p120 dynamically regulates Rho-GTPase activity at the cadherin complex through transient interaction with several of its up- and downstream effectors, including ROCK1. The American Society for Cell Biology 2012-01-01 /pmc/articles/PMC3248908/ /pubmed/22031287 http://dx.doi.org/10.1091/mbc.E11-06-0497 Text en © 2012 Smith 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 Smith, Andrew L. Dohn, Michael R. Brown, Meredith V. Reynolds, Albert B. Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin |
title | Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin |
title_full | Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin |
title_fullStr | Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin |
title_full_unstemmed | Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin |
title_short | Association of Rho-associated protein kinase 1 with E-cadherin complexes is mediated by p120-catenin |
title_sort | association of rho-associated protein kinase 1 with e-cadherin complexes is mediated by p120-catenin |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3248908/ https://www.ncbi.nlm.nih.gov/pubmed/22031287 http://dx.doi.org/10.1091/mbc.E11-06-0497 |
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