title: The cytoplasmic tyrosines of integrin subunit beta1 are involved in focal adhesion kinase activation.
abstract: We have previously shown that mutation of the two tyrosines in the cytoplasmic domain of integrin subunit beta1 (Y783 and Y795) to phenylalanines markedly reduces the capability of beta1A integrins to mediate directed cell migration. In this study, beta1-dependent cell spreading was found to be delayed in GD25 cells expressing beta1A(Y783/795F) compared to that in wild-type GD25-beta1A. Focal adhesion kinase (FAK) tyrosine phosphorylation and activation were severely impaired in response to beta1-dependent adhesion in GD25-beta1A(Y783/795F) cells compared to that in wild-type GD25-beta1A or mutants in which only a single tyrosine was altered (beta1A(Y783F) or beta1A(Y795F)). Phosphorylation site-specific antibodies selective for FAK phosphotyrosine 397 indicated that the defect in FAK phosphorylation via beta1A(Y783/795F) lies at the level of the initial autophosphorylation step. Indeed, beta1A-dependent tyrosine phosphorylation of tensin and paxillin was lost in the beta1A(Y783/795F) cells, consistent with the impairment in FAK activation. In contrast, p130(CAS) overall tyrosine phosphorylation was unaffected by the beta1 mutations. Despite the defect in beta1-mediated FAK activation, FAK was still localized to focal adhesions. Taken together, the phenotype of the GD25-beta1A(Y783/795F) cells resembles, but is distinct from, the phenotype observed in FAK-null cells. These observations argue that tyrosines 783 and 795 within the cytoplasmic tail of integrin subunit beta1A are critical mediators of FAK activation and cell spreading in GD25 cells.
authors.affiliation: Department of Medical Biochemistry and Microbiology, Uppsala University, S-751 23 Uppsala, Sweden. krister@med.unc.edu
authors.forename: K
authors.initials: K
authors.lastname: Wennerberg
authors.forename: A
authors.initials: A
authors.lastname: Armulik
authors.forename: T
authors.initials: T
authors.lastname: Sakai
authors.forename: M
authors.initials: M
authors.lastname: Karlsson
authors.forename: R
authors.initials: R
authors.lastname: Fässler
authors.forename: E M
authors.initials: EM
authors.lastname: Schaefer
authors.forename: D F
authors.initials: DF
authors.lastname: Mosher
authors.forename: S
authors.initials: S
authors.lastname: Johansson
mesh_terms: D019943:Amino Acid Substitution; D050719:Crk-Associated Substrate Protein; D003593:Cytoplasm / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004789:Enzyme Activation; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism*; D008840:Microfilament Proteins / Q000378:metabolism; D009154:Mutation; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011506:Proteins; D050717:Retinoblastoma-Like Protein p130; D015398:Signal Transduction; D000070876:Tensins; D014443:Tyrosine; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D050719:Crk-Associated Substrate Protein; D003598:Cytoskeletal Proteins; D019012:Integrin beta1; D008840:Microfilament Proteins; D051419:Paxillin; D010750:Phosphoproteins; D011506:Proteins; D050717:Retinoblastoma-Like Protein p130; D000070876:Tensins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases