Mechanisms of Signal Transduction
Association of the Tensin N-terminal Protein-tyrosine Phosphatase Domain with the α Isoform of Protein Phosphatase-1 in Focal Adhesions*

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Focal adhesions attach cultured cells to the extracellular matrix, and we found endogenous protein phosphatase-1α isoform (PP1α) localized in adhesions across the entire area of adherent fibroblasts. However, in fibroblasts migrating into a scrape wound or spreading after replating PP1α did not appear in adhesions near the leading edge but was recruited into other adhesions coincident in time and space with incorporation of tensin. Endogenous tensin and PP1α co-precipitated from cell lysates with isoform-specific PP1 antibodies. Chemical cross-linking of focal adhesion preparations with Lomant's reagent demonstrated molecular proximity of endogenous PP1α and tensin, whereas neither focal adhesion kinase nor vinculin was cross-linked and co-precipitated with PP1α, suggesting distinct spatial subdomains within adhesions. Transient expression of truncated tensin showed the N-terminal 360 residues, which comprise a protein-tyrosine phosphatase domain, alone were sufficient for isoform-selective co-precipitation of co-expressed PP1α. Human prostate cancer PC3 cells are deficient in tensin relative to fibroblasts and have fewer, mostly peripheral adhesions. Transient expression of green fluorescent protein tensin in these cancer cells induced formation of adhesions and recruited endogenous PP1α into those adhesions. Thus, the protein-tyrosine phosphatase domain of tensin exhibits isoform-specific association with PP1α in a restricted spatial region of adhesions that are formed during cell migration.

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This work was supported in part by grants from the United States Public Health Service, National Institutes of Health Grants CA 40042 (NCI) and Grant GM 56362 (NIGMS) and by a Scientist Development grant from the American Heart Association, National Center (to M. E.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.