Adhesion-mediated cytoskeletal remodeling is controlled by the direct scaffolding of Src from FAK complexes to lipid rafts by SSeCKS/AKAP12.
Metastatic cell migration and invasion are regulated by altered adhesion-mediated signaling to the actin-based cytoskeleton via activated Src-FAK complexes. Src-suppressed C-kinase substrate (SSeCKS, the rodent orthologue of human Gravin/AKAP12), whose expression is downregulated by oncogenic Src and in many human cancers, antagonizes oncogenic Src pathways including those driving neovascularization at metastatic sites, metastatic cell motility and invasiveness. This is likely manifested through its function as a scaffolder of F-actin and signaling proteins such as cyclins, calmodulin, protein kinase C and A. Here we show that in contrast to its ability to inhibit haptotaxis, SSeCKS increased prostate cancer cell adhesion to fibronectin and type I collagen in a FAK-dependent manner, correlating with a relative increase in FAK(poY397) levels. In contrast, SSeCKS suppressed adhesion-induced Src activation (Src(poY416)) and phosphorylation of FAK at Y925, a known Src substrate site. SSeCKS also induced increased cell spreading, cell flattening, integrin β1 clustering and formation of mature focal adhesion plaques. An in silico analysis identified a Src-binding domain on SSeCKS(aa 153-166) that is homologous to the Src-binding domain of caveolin-1, and this region is required for SSeCKS-Src interaction, for SSeCKS-enhanced Src activity and sequestration to lipid rafts and for SSeCKS-enhanced adhesion of MAT-LyLu and CWR22Rv1 prostate cancer cells. Our data suggest a model in which SSeCKS suppresses oncogenic motility by sequestering Src to caveolin-rich lipid rafts, thereby disengaging Src from FAK-associated adhesion and signaling complexes.
Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
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D054758:A Kinase Anchor Proteins / Q000235:genetics / Q000378:metabolism*; D008841:Actin Cytoskeleton / Q000378:metabolism*; D000595:Amino Acid Sequence; D051242:Caveolin 1 / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology; D002633:Chemotaxis / Q000502:physiology; D003599:Cytoskeleton / Q000378:metabolism; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism*; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D021962:Membrane Microdomains / Q000201:enzymology*; D008969:Molecular Sequence Data; D015398:Signal Transduction; D019061:src-Family Kinases / Q000378:metabolism
D054758:A Kinase Anchor Proteins; C491162:AKAP12 protein, human; D051242:Caveolin 1; D018797:Cell Cycle Proteins; D019012:Integrin beta1; D051417:Focal Adhesion Kinase 1; D019061:src-Family Kinases