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...

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Bibliographic Details
Published in:Molecular biology of the cell 2012-01, Vol.23 (1), p.99-110
Main Authors: Smith, Andrew L, Dohn, Michael R, Brown, Meredith V, Reynolds, Albert B
Format: Article
Language:English
Subjects:
Actins - metabolism
Cadherins - metabolism
Catenins - metabolism
Cell Line
Gene Knockdown Techniques
Guanine Nucleotide Exchange Factors - metabolism
Humans
Intercellular Junctions - metabolism
Microscopy, Fluorescence
Multiprotein Complexes - metabolism
Protein Binding
Protein Interaction Mapping
Protein Transport
Repressor Proteins - metabolism
rho-Associated Kinases - genetics
rho-Associated Kinases - metabolism
RNA Interference
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Summary: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.
ISSN:1059-1524
1939-4586
DOI:10.1091/mbc.E11-06-0497