title: Evidence that beta3 integrin-induced Rac activation involves the calpain-dependent formation of integrin clusters that are distinct from the focal complexes and focal adhesions that form as Rac and RhoA become active.
abstract: Interaction of integrins with the extracellular matrix leads to transmission of signals, cytoskeletal reorganizations, and changes in cell behavior. While many signaling molecules are known to be activated within Rac-induced focal complexes or Rho-induced focal adhesions, the way in which integrin-mediated adhesion leads to activation of Rac and Rho is not known. In the present study, we identified clusters of integrin that formed upstream of Rac activation. These clusters contained a Rac-binding protein(s) and appeared to be involved in Rac activation. The integrin clusters contained calpain and calpain-cleaved beta3 integrin, while the focal complexes and focal adhesions that formed once Rac and Rho were activated did not. Moreover, the integrin clusters were dependent on calpain for their formation. In contrast, while Rac- and Rho-GTPases were dependent on calpain for their activation, formation of focal complexes and focal adhesions by constitutively active Rac or Rho, respectively, occurred even when calpain inhibitors were present. Taken together, these data are consistent with a model in which integrin-induced Rac activation requires the formation of integrin clusters. The clusters form in a calpain-dependent manner, contain calpain, calpain-cleaved integrin, and a Rac binding protein(s). Once Rac is activated, other integrin signaling complexes are formed by a calpain-independent mechanism(s).
authors.affiliation: Joseph J. Jacobs Center for Thrombosis and Vascular Biology, Department of Molecular Cardiology, The Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
authors.forename: K
authors.initials: K
authors.lastname: Bialkowska
authors.forename: S
authors.initials: S
authors.lastname: Kulkarni
authors.forename: X
authors.initials: X
authors.lastname: Du
authors.forename: D E
authors.initials: DE
authors.lastname: Goll
authors.forename: T C
authors.initials: TC
authors.lastname: Saido
authors.forename: J E
authors.initials: JE
authors.lastname: Fox
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001011:Aorta; D002154:Calpain / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D002448:Cell Adhesion; D048429:Cell Size; D022081:Cell Surface Extensions / Q000378:metabolism; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D004789:Enzyme Activation; D005353:Fibronectins / Q000378:metabolism; D022001:Focal Adhesions / Q000737:chemistry / Q000378:metabolism*; D005799:Genes, Dominant / Q000235:genetics; D006801:Humans; D039661:Integrin beta3; D046911:Macromolecular Substances; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D016596:Vinculin / Q000378:metabolism; D019096:Vitronectin / Q000378:metabolism; D020744:rac GTP-Binding Proteins / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; D005353:Fibronectins; D039661:Integrin beta3; D046911:Macromolecular Substances; D010980:Platelet Membrane Glycoproteins; D019096:Vitronectin; D016596:Vinculin; D002154:Calpain; D020744:rac GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein