Pressure activates colon cancer cell adhesion via paxillin phosphorylation, Crk, Cas, and Rac1.
Physical forces can activate colon cancer cell adhesion, critical for metastasis. Paxillin is phosphorylated by FAK and required for pressure-stimulated adhesion. However, whether paxillin acts as an inert scaffolding protein or whether paxillin phosphorylation is required is unknown. Transfection with paxillin point-phosphorylation mutants demonstrated that phosphorylation at tyrosines 31 and 118 together is necessary for pressure-stimulated adhesion. We further evaluated potential paxillin partners. Reducing the adaptor protein Crk or the focal adhesion protein p130Cas blocked pressure-stimulated adhesion. Furthermore, Crk and p130Cas both displayed increased co-immunoprecipitation with paxillin in response to increased pressure, except in cells transfected with a Y31Y118 paxillin mutant. Inhibiting the small GTPase Rac1 also abolished pressure-stimulated adhesion, and reducing paxillin by siRNA blocked Rac1 phosphorylation by pressure. Thus, paxillin phosphorylation at tyrosines 31 and 118 together is necessary for pressure-induced adhesion. Paxillin, Crk and Cas form a trimeric complex that activates Rac1 and mediates this effect.
Downey
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C
Department of Surgery, John D. Dingell VA Medical Center and Wayne State University, 4646 John R. Street, Detroit, MI 48201, USA.
Craig
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DH
Basson
M D
MD
D000185:Actinin / Q000037:antagonists & inhibitors; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D050719:Crk-Associated Substrate Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D009362:Neoplasm Metastasis; D051419:Paxillin / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011312:Pressure; D050737:Proto-Oncogene Proteins c-crk / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034622:RNA Interference; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
D050719:Crk-Associated Substrate Protein; D051419:Paxillin; D050737:Proto-Oncogene Proteins c-crk; D000185:Actinin; D014443:Tyrosine; D020830:rac1 GTP-Binding Protein