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Required role of focal adhesion kinase (FAK) for integrin-stimulated cell migration
D.J. Sieg, C.R. Hauck, D.D. Schlaepfer
Journal of Cell Science 1999 112: 2677-2691;

Summary

FAK localizes to sites of transmembrane integrin receptor clustering and facilitates intracellular signaling events. FAK-null (FAK-) fibroblasts exhibit a rounded morphology, defects in cell migration, and an elevated number of cell-substratum contact sites. Here we show that stable re-expression of epitope-tagged FAK reversed the morphological defects of the FAK- cells through the dynamic regulation of actin structures and focal contact sites in fibronectin (FN) stimulated cells. FAK re-expressing fibroblasts (clones DA2 and DP3) exhibit a characteristic fibrillar shape and display indistinguishable FN receptor-stimulated migration properties compared to normal fibroblasts. Expression of various FAK mutants in the FAK- cells showed that FAK kinase activity, the Tyr-397/SH2 domain binding site, and the first proline-rich SH3 binding region in the FAK C-terminal domain were individually needed to promote full FAK-mediated FAK- cell migration to FN whereas direct paxillin binding to FAK was not required. Expression of the FAK Phe-397 mutant did not promote FAK- cell migration and overexpression of p50(csk) in DA2 cells inhibited migration to FN suggesting that Src-family PTKs play important roles in FAK-mediated motility events. Expression of the FAK C-terminal domain, FRNK, promoted FAK dephosphorylation at Tyr-397 and potently blocked FAK-mediated cell migration. This dominant-negative effect of FRNK was reversed by a point mutation (Leu-1034 to Ser) which prevented FRNK localization to focal contact sites. Our results show that FAK functions as a key regulator of fibronectin receptor stimulated cell migration events through the recruitment of both SH2 and SH3 domain-containing signaling proteins to sites of integrin receptor clustering.

REFERENCES

    1. Anand-Apte B.,
    2. Zetter B. R.,
    3. Viswanathan A.,
    4. Qiu R. G.,
    5. Chen J.,
    6. Ruggieri R. and
    7. Symons M.
    (1997). Platelet-derived growth factor and fibronectin-stimulated migration are differentially regulated by the Rac and extracellular signal-regulated kinase pathways. J. Biol. Chem 272, 30688–30692
    OpenUrlAbstract/FREE Full Text
    1. Angers-Loustau A.,
    2. Cote J.-F.,
    3. Charest A.,
    4. Dowbenko D.,
    5. Spencer S.,
    6. Lasky L. A. and
    7. Tremblay M. L.
    (1999). Protein tyrosine phosphatase-PEST regulates focal adhesion disassembly, migration, and cytokinesis in fibroblasts. J. Cell Biol 144, 1019–1031
    OpenUrlAbstract/FREE Full Text
    1. Calalb M.,
    2. Polte T. and
    3. Hanks S. K.
    (1995). Tyrosine phosphorylation of focal adhesion kinase at sites in the catalytic domain regulates kinase activity: a role for the Src family kinases. Mol. Cell. Biol 15, 954–963
    OpenUrlAbstract/FREE Full Text
    1. Cary L. A.,
    2. Chang J. F. and
    3. Guan J.-L.
    (1996). Stimulation of cell migration by overexpression of focal adhesion kinase and its association with Src and Fyn. J. Cell Sci 108, 1787–1794
    OpenUrl
    1. Cary L. A.,
    2. Han D. C.,
    3. Polte T. R.,
    4. Hanks S. K. and
    5. Guan J.-L.
    (1998). Identification of p130Casas a mediator of focal adhesion kinase-promoted cell migration. J. Cell Biol 140, 211–221
    OpenUrlAbstract/FREE Full Text
    1. Chen H. C.,
    2. Appeddu P. A.,
    3. Isoda H. and
    4. Guan J. L.
    (1996). Phosphorylation of tyrosine 397 in focal adhesion kinase is required for binding phosphatidylinositol 3-kinase. J. Biol. Chem 271, 26329–26334
    OpenUrlAbstract/FREE Full Text
    1. Chen H. C.,
    2. Appeddu P. A.,
    3. Parsons J. T.,
    4. Hildebrand J. D.,
    5. Schaller M. D. and
    6. Guan J. L.
    (1995). Interaction of focal adhesion kinase with cytoskeletal protein talin. J. Biol. Chem 270, 16995–16999
    OpenUrlAbstract/FREE Full Text
    1. Fincham V. J. and
    2. Frame M. C.
    (1998). The catalytic activity of Src is dispensable for translocation to focal adhesions but controls the turnover of these structures during cell motility. EMBO J 17, 81–92
    OpenUrlAbstract
    1. Frisch S. M.,
    2. Vuori K.,
    3. Ruoslahti E. and
    4. Chan-Hui P. Y.
    (1996). Control of adhesion-dependent cell survival by focal adhesion kinase. J. Cell Biol 134, 793–799
    OpenUrlAbstract/FREE Full Text
    1. Furuta Y.,
    2. Ilic D.,
    3. Kanazawa S.,
    4. Takeda N.,
    5. Yamamoto T. and
    6. Aizawa S.
    (1995). Mesodermal defect in late phase of gastrulation by a targeted mutation of focal adhesion kinase, FAK. Oncogene 11, 1989–1995
    OpenUrlPubMedWeb of Science
    1. George E. L.,
    2. Georges-Labouesse E. N.,
    3. Patel-King R. S.,
    4. Rayburn H. and
    5. Hynes R. O.
    (1993). Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin. Development 119, 1079–1091
    OpenUrlAbstract
    1. Gilmore A. P. and
    2. Romer L. H.
    (1996). Inhibition of focal adhesion kinase (FAK) signaling in focal adhesions decreases cell motility and proliferation. Mol. Biol. Cell 7, 1209–1224
    OpenUrlAbstract/FREE Full Text
    1. Hanks S. K. and
    2. Polte T. R.
    (1997). Signaling through focal adhesion kinase. BioEssays 19, 137–145
    OpenUrlCrossRefPubMedWeb of Science
    1. Hildebrand J. D.,
    2. Schaller M. D. and
    3. Parsons J. T.
    (1995). Paxillin, a tyrosine phosphorylated focal adhesion-associated protein binds to the carboxyl terminal domain of focal adhesion kinase. Mol. Biol. Cell 6, 637–647
    OpenUrlAbstract/FREE Full Text
    1. Hildebrand J. D.,
    2. Taylor J. M. and
    3. Parsons J. T.
    (1996). An SH3 domain-containing GTPase-activating protein for Rho and Cdc42 associates with focal adhesion kinase. Mol. Cell. Biol 16, 3169–3178
    OpenUrlAbstract/FREE Full Text
    1. Honda H.,
    2. Oda H.,
    3. Nakamoto T.,
    4. Honda Z.,
    5. Sakai R.,
    6. Suzuki T.,
    7. Saito T.,
    8. Nakamura K.,
    9. Nakao K.,
    10. Ishikawa T. and
    11. et al.
    (1998). Cardiovascular anomaly, impaired actin bundling and resistance to Src-induced transformation in mice lacking p130Cas. Nature Genet 19, 361–365
    OpenUrlCrossRefPubMedWeb of Science
    1. Ilic D.,
    2. Furuta Y.,
    3. Kanazawa S.,
    4. Takeda N.,
    5. Sobue K.,
    6. Nakatsuji N.,
    7. Nomura S.,
    8. Fujimoto J.,
    9. Okada M.,
    10. Yamamoto T. and
    11. et al.
    (1995). Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice. Nature 377, 539–544
    OpenUrlCrossRefPubMedWeb of Science
    1. Ilic D.,
    2. Almeida E. A.,
    3. Schlaepfer D. D.,
    4. Dazin P.,
    5. Aizawa S. and
    6. Damsky C. H.
    (1998). Extracellular matrix survival signals transduced by focal adheison kinase suppress p53-mediated apoptosis. J. Cell Biol 143, 547–560
    OpenUrlAbstract/FREE Full Text
    1. Keely P. J.,
    2. Westwick J. K.,
    3. Whitehead I. P.,
    4. Der C. J. and
    5. Parise L. V.
    (1997). Cdc42 and Rac1 induce integrin-mediated cell motility and invasiveness through PI(3)K. Nature 390, 632–636
    OpenUrlCrossRefPubMed
    1. Klemke R. L.,
    2. Cai S.,
    3. Giannini A. L.,
    4. Gallagher P. J.,
    5. de Lanerolle P. and
    6. Cheresh D. A.
    (1997). Regulation of cell motility by mitogen-activated protein kinase. J. Cell Biol 137, 481–492
    OpenUrlAbstract/FREE Full Text
    1. Klemke R. L.,
    2. Leng J.,
    3. Molander R.,
    4. Brooks P. C.,
    5. Vuori K. and
    6. Cheresh D. A.
    (1998). Cas/Crk coupling serves as a ‘molecular switch’ for induction of cell migration. J. Cell Biol 140, 961–972
    OpenUrlAbstract/FREE Full Text
    1. Klinghoffer R. A.,
    2. Sachsenmaier C.,
    3. Cooper J. A. and
    4. Soriano P.
    (1999). Src family kinases are required for integrin but not PDGFR signal transduction. EMBO J 18, 2459–2471
    OpenUrlAbstract
    1. Owens L. V.,
    2. Xu L. H.,
    3. Craven R. J.,
    4. Dent G. A.,
    5. Weiner T. M.,
    6. Kornberg L.,
    7. Liu E. T. and
    8. Cance W. G.
    (1995). Overexpression of the focal adhesion kinase (p125FAK) in invasive human tumors. Cancer Res 55, 2752–2755
    OpenUrlAbstract/FREE Full Text
    1. Polte T. R. and
    2. Hanks S. K.
    (1995). Interaction between focal adhesion kinase and Crk-associated tyrosine kinase substrate p130Cas. Proc. Nat. Acad. Sci. USA 92, 10678–10682
    OpenUrlAbstract/FREE Full Text
    1. Polte T. R. and
    2. Hanks S. K.
    (1997). Complexes of focal adhesion kinase (FAK) and Crk-associated substrate (p130Cas) are elevated in cytoskeleton-associated fractions following adhesion and Src transformation. Requirements for Src kinase activity and FAK proline-rich motifs. J. Biol. Chem 272, 5501–5509
    OpenUrlAbstract/FREE Full Text
    1. Ren X. D.,
    2. Kiosses W. B. and
    3. Schwartz M. A.
    (1999). Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton. EMBO J 18, 578–585
    OpenUrlAbstract
    1. Richardson A. and
    2. Parsons J. T.
    (1996). A mechanism for regulation of the adhesion-associated protein tyrosine kinase pp125FAK. Nature 380, 538–540
    OpenUrlCrossRefPubMedWeb of Science
    1. Richardson A.,
    2. Malik R. K.,
    3. Hildebrand J. D. and
    4. Parsons J. T.
    (1997). Inhibition of cell spreading by expression of the C-terminal domain of focal adhesion kinase (FAK) is rescued by coexpression of Src or catalytically inactive FAK: a role for paxillin tyrosine phosphorylation. Mol. Cell Biol 17, 6906–6914
    OpenUrlAbstract/FREE Full Text
    1. Schaller M. D.,
    2. Borgman C. A. and
    3. Parsons J. T.
    (1993). Autonomous expression of a noncatalytic domain of the focal adhesion-associated protein tyrosine kinase pp125FAK. Mol. Cell. Biol 13, 785–791
    OpenUrlAbstract/FREE Full Text
    1. Schaller M. D.,
    2. Otey C. A.,
    3. Hildebrand J. D. and
    4. Parsons J. T.
    (1995). Focal adhesion kinase and paxillin bind peptides mimickingintegrin cytoplasmic domains. J. Cell Biol 130, 1181–1187
    OpenUrlAbstract/FREE Full Text
    1. Schlaepfer D. D.,
    2. Hanks S. K.,
    3. Hunter T. and
    4. van der Geer P.
    (1994). Integrin-mediated signal transduction linked to Ras pathway by Grb2 binding to focal adhesion kinase. Nature 372, 786–791
    OpenUrlCrossRefPubMedWeb of Science
    1. Schlaepfer D. D. and
    2. Hunter T.
    (1996). Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases. Mol. Cell. Biol 16, 5623–5633
    OpenUrlAbstract/FREE Full Text
    1. Schlaepfer D. D. and
    2. Hunter T.
    (1997). FAK overexpression enhances Ras-dependent integrin signaling to ERK2/mitogen-activated protein kinase through interactions with and activation of c-Src. J. Biol. Chem 272, 13189–13195
    OpenUrlAbstract/FREE Full Text
    1. Schlaepfer D. D.,
    2. Broome M. A. and
    3. Hunter T.
    (1997). Fibronectin-stimulated signaling from a focal adhesion kinase-c-Src complex: involvement of the Grb2, p130Cas, and Nck adaptor proteins. Mol. Cell. Biol 17, 1702–1713
    OpenUrlAbstract/FREE Full Text
    1. Schlaepfer D. D.,
    2. Jones K. C. and
    3. Hunter T.
    (1998). Multiple Grb2-mediated integrin-stimulated signaling pathways to ERK2/mitogen-activated protein kinase: Summation of both c-Src and FAK-initiated tyrosine phosphorylation events. Mol. Cell. Biol 18, 2571–2585
    OpenUrlAbstract/FREE Full Text
    1. Schlaepfer D. D.,
    2. Hauck C. R. and
    3. Sieg D. J.
    (1999). Signaling through focal adhesion kinase. Prog. Biophys. Mol. Biol 71, 435–478
    OpenUrlCrossRefPubMedWeb of Science
    1. Sieg D. J.,
    2. Ilic D.,
    3. Jones K. C.,
    4. Damsky C. H.,
    5. Hunter T. and
    6. Schlaepfer D. D.
    (1998). Pyk2 and Src-family protein-tyrosine kinases compensate for the loss of FAK in fibronectin-stimulated signaling events but Pyk2 does not fully function to enhance FAKcell migration. EMBO J 17, 5933–5947
    OpenUrlAbstract
    1. Stephens L. E.,
    2. Sutherland A. E.,
    3. Klimanskaya I. V.,
    4. Andrieux A.,
    5. Meneses J.,
    6. Pedersen R. A. and
    7. Damsky C. H.
    (1995). Deletion of1 integrins in mice results in inner cell mass failure and peri-implantation lethality. Genes Dev 9, 1883–1895
    OpenUrlAbstract/FREE Full Text
    1. Tachibana K.,
    2. Sato T.,
    3. D'Avirro N. and
    4. Morimoto C.
    (1995). Direct association of pp125FAKwith paxillin, the focal adhesion-targeting mechanism of pp125FAK. J. Exp. Med 182, 1089–1100
    OpenUrlAbstract/FREE Full Text
    1. Tachibana K.,
    2. Urano T.,
    3. Fujita H.,
    4. Ohashi Y.,
    5. Kamiguchi K.,
    6. Iwata S.,
    7. Hirai H. and
    8. Morimoto C.
    (1997). Tyrosine phosphorylation of Crk-associated substrates by focal adhesion kinase. A putative mechanism for the integrin-mediated tyrosine phosphorylation of Crk-associated substrates. J. Biol. Chem 272, 29083–29090
    OpenUrlAbstract/FREE Full Text
    1. Tamura M.,
    2. Gu J.,
    3. Matsumoto K.,
    4. Aota S.,
    5. Parsons R. and
    6. Yamada K. M.
    (1998). Inhibition of cell migration, spreading, and focal adhesions by tumor suppressor PTEN. Science 280, 1614–1617
    OpenUrlAbstract/FREE Full Text
    1. Tamura M.,
    2. Gu J.,
    3. Takino T. and
    4. Yamada K. M.
    (1999). Tumor suppressor PTEN inhibition of cell invasion, migration, and growth: differential involvement of focal adhesion kinase and p130Cas. Cancer Res 59, 442–449
    OpenUrlAbstract/FREE Full Text
    1. Taylor J. M.,
    2. Macklem M. M. and
    3. Parsons J. T.
    (1999). Cytoskeletal changes induced by GRAF, the GTPase regulator associated with focal adhesion kinase, are mediated by Rho. J. Cell Sci 112, 231–242
    OpenUrlAbstract/FREE Full Text
    1. Vuori K.,
    2. Hirai H.,
    3. Aizawa S. and
    4. Ruoslahti E.
    (1996). Induction of p130Cassignaling complex formation upon integrin-mediated cell adhesion: a role for the Src family kinases. Mol. Cell. Biol 16, 2606–2613
    OpenUrlAbstract/FREE Full Text
    1. Yu D. H.,
    2. Qu C. K.,
    3. Henegariu O.,
    4. Lu X. and
    5. Feng G. S.
    (1998). Protein-tyrosine phosphatase Shp-2 regulates cell spreading, migration, and focal adhesion. J. Biol. Chem 273, 21125–21131
    OpenUrlAbstract/FREE Full Text
    1. Zhao J.-H.,
    2. Reiske H. and
    3. Guan J.-L.
    (1998). Regulation of the cell cycle by focal adhesion kinase. J. Cell Biol 143, 1997–2008
    OpenUrlAbstract/FREE Full Text
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Journal Articles
Required role of focal adhesion kinase (FAK) for integrin-stimulated cell migration
D.J. Sieg, C.R. Hauck, D.D. Schlaepfer
Journal of Cell Science 1999 112: 2677-2691;
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