title: Dynamic modulation of cytoskeletal proteins linking integrins to signaling complexes in spreading cells. Role of skelemin in initial integrin-induced spreading.
abstract: Recently we showed that signaling across beta3-integrin leads to activation of calpain and formation of integrin clusters that are involved in Rac activation. The subsequent activation of Rac and Rho leads to the formation of focal complexes and focal adhesions, respectively. The goal of the present study was to determine whether different proteins link the integrin to the cytoskeleton in the different complexes. We show that talin is present in focal adhesions but not in the calpain-induced clusters. alpha-Actinin colocalized with integrin at various sites, including the calpain-induced clusters. Skelemin, a protein shown recently to interact with beta1- and beta3-integrin in vitro, colocalized with integrin in calpain-induced clusters but was absent from focal adhesions. Cells transiently expressing skelemin C2 motifs, which contain the integrin binding site, failed to form integrin clusters or to spread on a substrate for beta1- and beta3-integrins. These results 1) suggest a dynamic reorganization of integrin complexes during cell spreading, 2) show that different cytoskeletal proteins link integrins in different complexes, and 3) demonstrate that skelemin is responsible for linking integrin to the calpain-induced clusters, and 4) show that the integrin-skelemin interaction is essential for transmission of signals leading to the initial steps of cell spreading.
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 B
authors.initials: KB
authors.lastname: Reddy
authors.forename: K
authors.initials: K
authors.lastname: Bialkowska
authors.forename: J E
authors.initials: JE
authors.lastname: Fox
mesh_terms: D000185:Actinin / Q000737:chemistry / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D016466:CHO Cells; D002154:Calpain / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D064211:Connectin; D006224:Cricetinae; D003598:Cytoskeletal Proteins / Q000378:metabolism* / Q000502:physiology*; D003599:Cytoskeleton / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology; D004789:Enzyme Activation; D005347:Fibroblasts / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D022001:Focal Adhesions; D049452:Green Fluorescent Proteins; D006801:Humans; D039661:Integrin beta3; D016023:Integrins / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009124:Muscle Proteins; D010957:Plasmids / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D051381:Rats; D015398:Signal Transduction; D016608:Talin / Q000378:metabolism; D014162:Transfection
chemical_list: D015703:Antigens, CD; D064211:Connectin; D003598:Cytoskeletal Proteins; D005353:Fibronectins; D039661:Integrin beta3; D016023:Integrins; D008164:Luminescent Proteins; C577299:MYOM1 protein, human; D009124:Muscle Proteins; C577300:Myom1 protein, mouse; D010980:Platelet Membrane Glycoproteins; D016608:Talin; D000185:Actinin; D049452:Green Fluorescent Proteins; D002154:Calpain