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Journal of Cell Science 114, 2279-2289 (2001)
© 2001 The Company of Biologists Limited


RESEARCH ARTICLE

pp60c-src and related tyrosine kinases: a role in the assembly and reorganization of matrix adhesions

Tova Volberg1, Lewis Romer2,*, Eli Zamir1 and Benjamin Geiger1,{ddagger}

1 Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel
2 Departments of Pediatrics, Cell Biology and Anatomy, and Anesthesiology, University of North Carolina at Chapel Hill, NC 27599-7220, USA
* Present address: Departments of Anesthesiology and Cell Biology, Johns Hopkins University, Blalock 904, Baltimore, MD 21215-4904, USA

{ddagger}Author for correspondence (e-mail: benny.geiger{at}weizmann.ac.il)

Accepted March 20, 2001

Activation of tyrosine kinases during integrin-mediated cell-matrix adhesion is involved both in the regulation of focal contact assembly and in the initiation of signaling processes at the cell-matrix adhesive interface. In order to determine the role of pp60c-src and related kinases in these processes, we have compared the dynamic reorganization of phosphotyrosine, vinculin, focal adhesion kinase and tensin in cells with altered expression of Src-family kinases. Both null cells for pp60c-src and triple knockout cells for pp60c-src, pp59fyn, and pp62c-yes exhibited decreased phosphotyrosine levels in focal contacts when compared with wild-type cells. pp60c-src-null cells also exhibited faster assembly of cell-matrix adhesions and a more exuberant recruitment of FAK to these sites. Tensin, which normally segregates into fibrillar adhesions was localized in large focal contacts in the two mutant cell lines, suggesting involvement of pp60c-src in the segregation of focal contacts and fibrillar adhesions. Moreover, treatment of wild-type cells with tyrphostin AG1007, which inhibits both pp60c-src and FAK activity, induced accumulation of tensin in peripheral focal adhesions. These findings demonstrate that Src family kinases, and pp60c-src in particular, have a central role in regulating protein dynamics at cell-matrix interfaces, both during early stages of interaction and in mature focal contacts.

Key words: pp60c-src, Tensin, Focal contacts, Fibrillar adhesions, Cell-matrix adhesions, Tyrosine phosphorylation




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© The Company of Biologists Ltd 2001