Myosin II activity dependent and independent vinculin recruitment to the sites of E-cadherin-mediated cell-cell adhesion

Grant M. Sumida, Tyler M. Tomita, Wenting Shih, Soichiro Yamada

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Background: Maintaining proper adhesion between neighboring cells depends on the ability of cells to mechanically respond to tension at cell-cell junctions through the actin cytoskeleton. Thus, identifying the molecules involved in responding to cell tension would provide insight into the maintenance, regulation, and breakdown of cell-cell junctions during various biological processes. Vinculin, an actin-binding protein that associates with the cadherin complex, is recruited to cell-cell contacts under increased tension in a myosin II-dependent manner. However, the precise role of vinculin at force-bearing cell-cell junctions and how myosin II activity alters the recruitment of vinculin at quiescent cell-cell contacts have not been demonstrated.Results: We generated vinculin knockdown cells using shRNA specific to vinculin and MDCK epithelial cells. These vinculin-deficient MDCK cells form smaller cell clusters in a suspension than wild-type cells. In wound healing assays, GFP-vinculin accumulated at cell-cell junctions along the wound edge while vinculin-deficient cells displayed a slower wound closure rate compared to vinculin-expressing cells. In the presence of blebbistatin (myosin II inhibitor), vinculin localization at quiescent cell-cell contacts was unaffected while in the presence of jasplakinolide (F-actin stabilizer), vinculin recruitment increased in mature MDCK cell monolayers.Conclusion: These results demonstrate that vinculin plays an active role at adherens junctions under increased tension at cell-cell contacts where vinculin recruitment occurs in a myosin II activity-dependent manner, whereas vinculin recruitment to the quiescent cell-cell junctions depends on F-actin stabilization.

Original languageEnglish (US)
Article number48
JournalBMC Cell Biology
Volume12
DOIs
StatePublished - Nov 3 2011

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Myosin Type II
Vinculin
Cadherins
Cell Adhesion
Intercellular Junctions
Madin Darby Canine Kidney Cells
jasplakinolide
Actins

ASJC Scopus subject areas

  • Cell Biology

Cite this

Myosin II activity dependent and independent vinculin recruitment to the sites of E-cadherin-mediated cell-cell adhesion. / Sumida, Grant M.; Tomita, Tyler M.; Shih, Wenting; Yamada, Soichiro.

In: BMC Cell Biology, Vol. 12, 48, 03.11.2011.

Research output: Contribution to journalArticle

Sumida, Grant M. ; Tomita, Tyler M. ; Shih, Wenting ; Yamada, Soichiro. / Myosin II activity dependent and independent vinculin recruitment to the sites of E-cadherin-mediated cell-cell adhesion. In: BMC Cell Biology. 2011 ; Vol. 12.
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