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Volume 272, Number 51, Issue of December 19, 1997
pp. 32448-32453
(Received for publication, June 27, 1997, and in revised form, October 2, 1997)
From the Cadherins mediate calcium-dependent
cell-cell adhesion, and this activity is regulated by cytoplasmic
interactions between cadherins, catenins, and the actin-based
cytoskeleton.
Vinculin Is Associated with the E-cadherin Adhesion Complex
,
,
and
Department of Surgery, Memorial
Sloan-Kettering Cancer Center, New York, New York 10021 and the
¶ Department of Pathology, Yale University School of Medicine,
New Haven, Connecticut 06510
-Catenin plays a critical role in the transmembrane
anchorage of cadherins, and deletion of -catenin has been shown to
inactivate cadherin-mediated adhesion, resulting in a nonadhesive
phenotype. Here we show that serum starvation increases E-cadherin
expression and induces E-cadherin-dependent adhesion in the
MDA-MB-468 breast cancer cell line. This adhesion occurred despite a
lack of -catenin expression, which was caused by mutations in the
-catenin gene. Coprecipitation analysis suggests that this adhesion
may be mediated by cytoplasmic connections from cadherins to the
cytoskeleton involving vinculin. A high level of vinculin associated
with E-cadherin immunoprecipitates was observed in MDA-MB-468 cells. In
contrast, vinculin was not detected in E-cadherin complexes in the A431
and MCF-7 epithelial carcinoma cell lines, which express -catenin.
However, in reciprocal immunoprecipitations using anti-vinculin
antibodies, E-cadherin associated strongly with vinculin in MDA-MB-468
cells and, to a lesser extent, in A431 and MCF-7 cells. These results
suggest that both -catenin and vinculin may be present in the
adhesion complex. To test the hypothesis that vinculin associates with E-cadherin complexes via 
-catenin, excess recombinant -catenin or
-catenin fusion protein was added to MDA-MB-468 cell lysates. Both
specifically inhibited the coprecipitation of E-cadherin with vinculin,
suggesting competition for the same binding site. These results suggest
that vinculin plays a role in the establishment or regulation of the
cadherin-based cell adhesion complex by direct interaction with
-catenin.
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