The catalytic activity of Src is dispensable for translocation to focal adhesions but controls the turnover of these structures during cell motility.
The Src family of protein tyrosine kinases is involved in transducing signals at sites of cellular adhesion. In particular, the v-Src oncoprotein resides in cellular focal adhesions, where it induces tyrosine phosphorylation of pp125FAK and focal adhesion loss during transformation. v-Src is translocated to cellular focal adhesions by an actin-dependent process. Here we have used mutant v-Src proteins that are temperature-dependent for translocation, but with secondary mutations that render them constitutively kinase-inactive or myristylation-defective, to show that neither v-Src kinase activity nor a myristyl group are required to induce association of v-Src with actin stress fibres and redistribution to sites of focal adhesions at the stress fibre termini. Moreover, switching the constitutively kinase-inactive or myristylation-defective temperature-sensitive v-Src proteins to the permissive temperature resulted in concomitant association with tyrosine-phosphorylated focal adhesion kinase (pp125FAK) and redistribution of both to focal adhesions. However, both catalytic activity and myristylation-mediated membrane association are required to induce dissociation of pp125FAK from v-Src, later degradation of pp125FAK and focal adhesion turnover during transformation and cell motility. These observations provide strong evidence that the role of the tyrosine kinase activity of the Src family at sites of cellular focal adhesions is to regulate the turnover of these structures during cell motility.
Fincham
V J
VJ
The Beatson Institute for Cancer Research, CRC Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD, UK.
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M C
MC
D000818:Animals; D001692:Biological Transport; D002384:Catalysis; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002465:Cell Movement; D002478:Cells, Cultured; D002642:Chick Embryo; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006868:Hydrolysis; D019814:Myristic Acid / Q000378:metabolism; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013696:Temperature; D019061:src-Family Kinases / Q000378:metabolism
D015815:Cell Adhesion Molecules; D019814:Myristic Acid; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src); D019061:src-Family Kinases