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Integrin-linked kinase is localized to cell-matrix focal adhesions but not cell-cell adhesion sites and the focal adhesion localization of integrin-linked kinase is regulated by the PINCH-binding ANK repeats
F. Li, Y. Zhang, C. Wu
Journal of Cell Science 1999 112: 4589-4599;

Summary

Integrin-linked kinase (ILK) is a ubiquitously expressed protein serine/threonine kinase that has been implicated in integrin-, growth factor- and Wnt-signaling pathways. In this study, we show that ILK is a constituent of cell-matrix focal adhesions. ILK was recruited to focal adhesions in all types of cells examined upon adhesion to a variety of extracellular matrix proteins. By contrast, ILK was absent in E-cadherin-mediated cell-cell adherens junctions. In previous studies, we have identified PINCH, a protein consisting of five LIM domains, as an ILK binding protein. We demonstrate in this study that the ILK-PINCH interaction requires the N-terminal-most ANK repeat (ANK1) of ILK and one (the C-terminal) of the two zinc-binding modules within the LIM1 domain of PINCH. The ILK ANK repeats domain, which is capable of interacting with PINCH in vitro, could also form a complex with PINCH in vivo. However, the efficiency of the complex formation or the stability of the complex was markedly reduced in the absence of the C-terminal domain of ILK. The PINCH binding defective ANK1 deletion ILK mutant, unlike the wild-type ILK, was unable to localize and cluster in focal adhesions, suggesting that the interaction with PINCH is necessary for focal adhesion localization and clustering of ILK. The N-terminal ANK repeats domain, however, is not sufficient for mediating focal adhesion localization of ILK, as an ILK mutant containing the ANK repeats domain but lacking the C-terminal integrin binding site failed to localize in focal adhesions. These results suggest that focal adhesions are a major subcellular compartment where ILK functions in intracellular signal transduction, and provide important evidence for a critical role of PINCH and integrins in regulating ILK cellular function.

REFERENCES

    1. Barreuther, M. F. and
    2. Grabel, L. B.
    (1996). The role of phosphorylation in modulating beta 1 integrin localization. Exp. Cell Res 222, 10–15.
    OpenUrlCrossRefPubMed
    1. Barry, S. T. and
    2. Critchley, D. R.
    (1994). The RhoA-dependent assembly of focal adhesions in Swiss 3T3 cells is associated with increased tyrosine phosphorylation and the recruitment of both pp125FAK and protein kinase C-delta to focal adhesions. J. Cell Sci 107, 2033–2045.
    OpenUrlAbstract/FREE Full Text
    1. Barstead, R. J. and
    2. Waterston, R. H.
    (1991). Vinculin is essential for muscle function in the nematode. J. Cell Biol 114, 715–724.
    OpenUrlAbstract/FREE Full Text
    1. Beckerle, M. C.
    (1990). The adhesion plaque protein, talin, is phosphorylated in vivo in chicken embryo fibroblasts exposed to a tumor-promoting phorbol ester. Cell Reg 1, 227–236.
    OpenUrlPubMedWeb of Science
    1. Beckerle, M. C.
    (1997). Zyxin: zinc fingers at sites of cell adhesion. [Review] [71 refs]. BioEssays 19, 949–957.
    OpenUrlCrossRefPubMedWeb of Science
    1. Bellis, S. L.,
    2. Perrotta, J. A.,
    3. Curtis, M. S. and
    4. Turner, C. E.
    (1997). Adhesion of fibroblasts to fibronectin stimulates both serine and tyrosine phosphorylation of paxillin. Biochem. J 325, 375–381.
    OpenUrlAbstract/FREE Full Text
    1. Bishop, L. A.,
    2. Kee, W. J.,
    3. Zhu, A. J. and
    4. Watt, F. M.
    (1998). Lack of intrinsic polarity in the ligand-binding ability of keratinocyte beta1 integrins. Exp. Dermatol 7, 350–361.
    OpenUrlCrossRefPubMedWeb of Science
    1. Braga, V. M.,
    2. Hajibagheri, N. and
    3. Watt, F. M.
    (1998). Calcium-induced intercellular adhesion of keratinocytes does not involve accumulation of beta 1 integrins at cell-cell contacts and does not involve changes in the levels or phosphorylation of catenins. Cell Adhes. Commun 5, 137–149.
    OpenUrlCrossRefPubMed
    1. Brown, M. C.,
    2. Perrotta, J. A. and
    3. Turner, C. E.
    (1996). Identification of LIM3 as the principal determinant of paxillin focal adhesion localization and characterization of a novel motif on paxillin directing vinculin and focal adhesion kinase binding. J. Cell Biol 135, 1109–1123.
    OpenUrlAbstract/FREE Full Text
    1. Brown, M. C.,
    2. Perrotta, J. A. and
    3. Turner, C. E.
    (1998). Serine and threonine phosphorylation of the paxillin LIM domains regulates paxillin focal adhesion localization and cell adhesion to fibronectin. Mol. Biol. Cell 9, 1803–1816.
    OpenUrlAbstract/FREE Full Text
    1. Burridge, K. and
    2. Chrzanowska-Wodnicka, M.
    (1996). Focal adhesions, contractility and signaling. Annu. Rev. Cell Dev. Biol 12, 463–518.
    OpenUrlCrossRefPubMedWeb of Science
    1. Chung, D. H.,
    2. Lee, J. I.,
    3. Kook, M. C.,
    4. Kim, J. R.,
    5. Kim, S. H.,
    6. Choi, E. Y.,
    7. Park, S. H. and
    8. Song, H. G.
    (1998). ILK (beta1-integrin-linked protein kinase): a novel immunohistochemical marker for Ewing's sarcoma and primitive neuroectodermal tumour. Virchows Archiv 433, 113–117.
    OpenUrlCrossRefPubMed
    1. De Nichilo, M. O. and
    2. Yamada, K. M.
    (1996). Integrin alpha v beta 5-dependent serine phosphorylation of paxillin in cultured human macrophages adherent to vitronectin. J. Biol. Chem 271, 11016–11022.
    OpenUrlAbstract/FREE Full Text
    1. Dedhar, S. and
    2. Hannigan, G. E.
    (1996). Integrin cytoplasmic interactions and bidirectional transmembrane signalling. [Review] [86 refs]. Curr. Opin. Cell Biol 8, 657–669.
    OpenUrlCrossRefPubMedWeb of Science
    1. Delcommenne, M.,
    2. Tan, C.,
    3. Gray, V.,
    4. Rue, L.,
    5. Woodgett, J. and
    6. Dedhar, S.
    (1998). Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. Proc. Nat. Acad. Sci. USA 95, 11211–11216.
    OpenUrlAbstract/FREE Full Text
    1. Freed, E.,
    2. Gailit, J.,
    3. van der Geer, P.,
    4. Ruoslahti, E. and
    5. Hunter, T.
    (1989). A novel integrin beta subunit is associated with the vitronectin receptor alpha subunit (alpha v) in a human osteosarcoma cell line and is a substrate for protein kinase C. EMBO J 8, 2955–2965.
    OpenUrlPubMedWeb of Science
    1. Gettner, S. N.,
    2. Kenyon, C. and
    3. Reichardt, L. F.
    (1995). Characterization ofpat-3 heterodimers, a family of essential integrin receptors in C. elegans. J. Cell Biol 129, 1127–1141.
    OpenUrlAbstract/FREE Full Text
    1. Hannigan, G. E.,
    2. Leung-Hagesteijn, C.,
    3. Fitz-Gibbon, L.,
    4. Coppolino, M. G.,
    5. Radeva, G.,
    6. Filmus, J.,
    7. Bell, J. C. and
    8. Dedhar, S.
    (1996). Regulation of cell adhesion and anchorage-dependent growth by a new beta 1-integrin-linked protein kinase. Nature 379, 91–96.
    OpenUrlCrossRefPubMed
    1. Hobert, O.,
    2. Moerman, D. G.,
    3. Clark, K. A.,
    4. Beckerle, M. C. and
    5. Ruvkun, G.
    (1999). A conserved LIM protein that affects muscular adherens junction integrity and mechanosensory function in Caenorhabditis elegans. J. Cell Biol 144, 45–57.
    OpenUrlAbstract/FREE Full Text
    1. Hogervorst, F.,
    2. Kuikman, I.,
    3. Noteboom, E. and
    4. Sonnenberg, A.
    (1993). The role of phosphorylation in activation of the alpha 6A beta 1 laminin receptor. J. Biol. Chem 268, 18427–18430.
    OpenUrlAbstract/FREE Full Text
    1. Ilic, D.,
    2. Damsky, C. H. and
    3. Yamamoto, T.
    (1997). Focal adhesion kinase: at the crossroads of signal transduction. J. Cell Sci 110, 401–407.
    OpenUrlAbstract/FREE Full Text
    1. Jaken, S.,
    2. Leach, K. and
    3. Klauck, T.
    (1989). Association of type 3 protein kinase C with focal contacts in rat embryo fibroblasts. J. Cell Biol 109, 697–704.
    OpenUrlAbstract/FREE Full Text
    1. Jockusch, B. M.,
    2. Bubeck, P.,
    3. Giehl, K.,
    4. Kroemker, M.,
    5. Moschner, J.,
    6. Rothkegel, M.,
    7. Rudiger, M.,
    8. Schluter, K.,
    9. Stanke, G. and
    10. Winkler, J.
    (1995). The molecular architecture of focal adhesions. [Review] [291 refs]. Annu. Rev. Cell Dev. Biol 11, 379–416.
    OpenUrlCrossRefPubMedWeb of Science
    1. Kim, L. T. and
    2. Yamada, K. M.
    (1997). Evidence that beta1 integrins in keratinocyte cell-cell junctions are not in the ligand-occupied conformation. J. Inv. Dermatol 108, 876–880.
    OpenUrlCrossRefPubMed
    1. LaFlamme, S. E.,
    2. Thomas, L. A.,
    3. Yamada, S. S. and
    4. Yamada, K. M.
    (1994). Single subunit chimeric integrins as mimics and inhibitors of endogenous integrin functions in receptor localization, cell spreading and migration and matrix assembly. J. Cell Biol 126, 1287–1298.
    OpenUrlAbstract/FREE Full Text
    1. Li, F.,
    2. Liu, J.,
    3. Mayne, R. and
    4. Wu, C.
    (1997). Identification and characterization of a mouse protein kinase that is highly homologous to human integrin-linked kinase. Biochim. Biophys. Acta 1358, 215–220.
    OpenUrlPubMed
    1. Novak, A.,
    2. Hsu, S. C.,
    3. Leung-Hagesteijn, C.,
    4. Radeva, G.,
    5. Papkoff, J.,
    6. Montesano, R.,
    7. Roskelley, C.,
    8. Grosschedl, R. and
    9. Dedhar, S.
    (1998). Cell adhesion and the integrin-linked kinase regulate the LEF-1 and beta-catenin signaling pathways. Proc. Nat. Acad. Sci. USA 95, 4374–4379.
    OpenUrlAbstract/FREE Full Text
    1. Oh, E. S.,
    2. Woods, A. and
    3. Couchman, J. R.
    (1997). Syndecan-4 proteoglycan regulates the distribution and activity of protein kinase C. J. Biol. Chem 272, 8133–8136.
    OpenUrlAbstract/FREE Full Text
    1. Oh, E. S.,
    2. Woods, A. and
    3. Couchman, J. R.
    (1997). Multimerization of the cytoplasmic domain of syndecan-4 is required for its ability to activate protein kinase C. J. Biol. Chem 272, 11805–11811.
    OpenUrlAbstract/FREE Full Text
    1. Parsons, J. T. and
    2. Parsons, S. J.
    (1997). Src family protein tyrosine kinases: cooperating with growth factor and adhesion signaling pathways. [Review] [56 refs]. Curr. Opin. Cell Biol 9, 187–192.
    OpenUrlCrossRefPubMedWeb of Science
    1. Pavalko, F. M.,
    2. Adam, L. P.,
    3. Wu, M. F.,
    4. Walker, T. L. and
    5. Gunst, S. J.
    (1995). Phosphorylation of dense-plaque proteins talin and paxillin during tracheal smooth muscle contraction. Am. J. Physiol 268, 563–571.
    OpenUrl
    1. Radeva, G.,
    2. Petrocelli, T.,
    3. Behrend, E.,
    4. Leung-Hagesteijn, C.,
    5. Filmus, J.,
    6. Slingerland, J. and
    7. Dedhar, S.
    (1997). Overexpression of the integrin-linked kinase promotes anchorage-independent cell cycle progression. J. Biol. Chem 272, 13937–13944.
    OpenUrlAbstract/FREE Full Text
    1. Reszka, A. A.,
    2. Hayashi, Y. and
    3. Horwitz, A. F.
    (1992). Identification of amino acid sequences in the integrin beta 1 cytoplasmic domain implicated in cytoskeletal association. J. Cell Biol 117, 1321–1330.
    OpenUrlAbstract/FREE Full Text
    1. Richardson, A.,
    2. Shannon, J. D.,
    3. Adams, R. B.,
    4. Schaller, M. D. and
    5. Parsons, J.
    (1997). Identification of integrin-stimulated sites of serine phosphorylation in FRNK, the separately expressed C-terminal domain of focal adhesion kinase: a potential role for protein kinase A. Biochem. J 324, 141–149.
    OpenUrlAbstract/FREE Full Text
    1. Rogalski, T. M.,
    2. Williams, B. D.,
    3. Mullen, G. P. and
    4. Moerman, D. G.
    (1993). Products of the unc-52 gene in Caenorhabditis elegans are homologous to the core protein of the mammalian basement membrane heparan sulfate proteoglycan. Genes Dev 7, 1471–1484.
    OpenUrlAbstract/FREE Full Text
    1. Roman, J.,
    2. LaChance, R. M.,
    3. Broekelmann, T. J.,
    4. Kennedy, C. J.,
    5. Wayner, E. A.,
    6. Carter, W. G. and
    7. McDonald, J. A.
    (1989). The fibronectin receptor is organized by extracellular matrix fibronectin: implications for oncogenic transformation and for cell recognition of fibronectin matrices. J. Cell Biol 108, 2529–2543.
    OpenUrlAbstract/FREE Full Text
    1. Sadler, I.,
    2. Crawford, A. W.,
    3. Michelsen, J. W. and
    4. Beckerle, M. C.
    (1992). Zyxin and cCRP: two interactive LIM domain proteins associated with the cytoskeleton. J. Cell Biol 119, 1573–1587.
    OpenUrlAbstract/FREE Full Text
    1. Schmeichel, K. L. and
    2. Beckerle, M. C.
    (1997). Molecular dissection of a LIM domain. Mol. Biol. Cell 8, 219–230.
    OpenUrlAbstract/FREE Full Text
    1. Schwartz, M. A.,
    2. Schaller, M. D. and
    3. Ginsberg, M. H.
    (1995). Integrins: emerging paradigms of signal transduction. Annu. Rev. Cell Dev. Biol 11, 549–599.
    OpenUrlCrossRefPubMedWeb of Science
    1. Tachibana, K.,
    2. Sato, T.,
    3. N, D. A. and
    4. Morimoto, C.
    (1995). Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. J. Exp. Med 182, 1089–1099.
    OpenUrlAbstract/FREE Full Text
    1. Tu, Y.,
    2. Li, F. and
    3. Wu, C.
    (1998). Nck-2, a novel Src homology2/3-containing adaptor protein that interacts with the LIM-only protein PINCH and components of growth factor receptor kinase signaling pathways. Mol. Biol. Cell 9, 3367–3382.
    OpenUrlAbstract/FREE Full Text
    1. Tu, Y.,
    2. Li, F.,
    3. Goicoechea, S. and
    4. Wu, C.
    (1999). The LIM-only protein PINCH directly interacts with integrin-linked kinase and is recruited to integrin-rich sites in spreading cells. Mol. Cell. Biol 19, 2425–2434.
    OpenUrlAbstract/FREE Full Text
    1. Turner, C. E.,
    2. Glenney, J. R. Jr.. and
    3. Burridge, K.
    (1990). Paxillin: a new vinculin-binding protein present in focal adhesions. J. Cell Biol 111, 1059–1068.
    OpenUrlAbstract/FREE Full Text
    1. Woods, A. and
    2. Couchman, J. R.
    (1992). Protein kinase C involvement in focal adhesion formation. J. Cell Sci 101, 277–290.
    OpenUrlAbstract/FREE Full Text
    1. Wu, C.,
    2. Keightley, S. Y.,
    3. Leung-Hagesteijn, C.,
    4. Radeva, G.,
    5. Coppolino, M.,
    6. Goicoechea, S.,
    7. McDonald, J. A. and
    8. Dedhar, S.
    (1998). Integrin-linked protein kinase regulates fibronectin matrix assembly, E-cadherin expression and tumorigenicity. J. Biol. Chem 273, 528–536.
    OpenUrlAbstract/FREE Full Text
    1. Xie, W.,
    2. Li, F.,
    3. Kudlow, J. E. and
    4. Wu, C.
    (1998). Expression of the integrin-linked kinase (ILK) in mouse skin: loss of expression in suprabasal layers of the epidermis and up-regulation by erbB-2. Am. J. Pathol 153, 367–372.
    OpenUrlCrossRefPubMed
    1. Yamakita, Y.,
    2. Totsukawa, G.,
    3. Yamashiro, S.,
    4. Fry, D.,
    5. Zhang, X.,
    6. Hanks, S. K. and
    7. Matsumura, F.
    (1999). Dissociation of FAK/p130(CAS)/c-Src complex during mitosis: role of mitosis-specific serine phosphorylation of FAK. J. Cell Biol 144, 315–324.
    OpenUrlAbstract/FREE Full Text
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Journal Article
Integrin-linked kinase is localized to cell-matrix focal adhesions but not cell-cell adhesion sites and the focal adhesion localization of integrin-linked kinase is regulated by the PINCH-binding ANK repeats
F. Li, Y. Zhang, C. Wu
Journal of Cell Science 1999 112: 4589-4599;
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