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. 2004 Jun;24(3):461–475. doi: 10.1023/B:CEMN.0000022774.72027.0e

Nerve Growth Factor (NGF) Induces a Rapid and Sustained Downregulation of the Focal Adhesion Kinase (FAK)

A Gatti 1
PMCID: PMC11529940  PMID: 15206825

Abstract

1. Exposure of PC12 cells to nerve growth factor (NGF) induces an early tyrosine phosphorylation of many proteins, a number of which is still unidentified. Although NGF is known to bind to and activate the receptor tyrosine kinase TrkA, many downstream targets of NGF signaling may be possibly phosphorylated by nonreceptor tyrosine kinases such as c-Src and focal adhesion kinase (FAK).

2. In the present study, exposure of TrkA-overexpressing PC12 cells to NGF is found to cause a rapid and sustained loss in the recovery of a subpopulation of nominally active FAK (i.e., being autophosphorylated on the positive site of regulation).

3. Consistent with the possibility that NGF induces the proteolysis of FAK via recruitment of Src family kinases, the use of various phosphorylation site-specific anti-FAK antibodies revealed an NGF-inducible and PP1-sensitive accumulation of a putative fragment (i.e., p62) of FAK. Significantly, the mitogenic epidermal growth factor (EGF) failed to induce the downregulation of FAK and the accumulation of tyrosine phosphorylated p62. Such differential response of FAK to NGF and EGF may shape the specificity by which these growth factors control the status of cell–matrix adhesion and the adhesion-driven signaling.

Keywords: adhesion, epidermal growth factor, focal adhesion kinase, nerve growth factor, PC12 cells, Src family kinases

REFERENCES

  1. Abedi, H., Dawes, K. E., and Zachary, I. (1995). Differential effects of platelet-derived growth factor BB on p125 focal adhesion kinase and paxillin tyrosine phosphorylation and on cell migration in rabbit aortic vascular smooth muscle cells and Swiss 3T3 fibroblasts. J. Biol. Chem.270:11367-11376. [DOI] [PubMed] [Google Scholar]
  2. Alema', S., Casalbore, P., Agostini, E., and Tato', F. (1985). Differentiation of PC12 pheochromocytoma cells induced by v-src oncogene. Nature316:557-559. [DOI] [PubMed] [Google Scholar]
  3. Baron, V., Calleja, V., Ferrari, P., Alengrin, F., and Van Obberghen, E. (1998). p125 Fak focal adhesion kinase is a substrate for the insulin and insulin-like growth factor-I tyrosine kinase receptors. J. Biol. Chem.273:7162-7168. [DOI] [PubMed] [Google Scholar]
  4. Calalb, M. B., Polte, T. R., and Hanks, S. K. (1995). Tyrosine phosphorylation of focal adhesion kinase at sites in the catalytic domain regulates kinase activity: A role for Src family kinases. Mol. Cell. Biol.15:954-963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Calalb, M. B., Zhang, X., Polte, T. R., and Hanks, S. K. (1996). Focal adhesion kinase tyrosine-861 is a major site of phosphorylation by Src. Biochem. Biophys. Res. Commun.228:662-668. [DOI] [PubMed] [Google Scholar]
  6. Carragher, N. O., Fincham, V. J., Riley, D., and Frame, M. C. (2001). Cleavage of focal adhesion kinase by different proteases during SRC-regulated transformation and apoptosis. Distinct roles for calpain and caspases. J. Biol. Chem.276:4270-4275. [DOI] [PubMed] [Google Scholar]
  7. Casamassima, A., and Rozengurt, E. (1997). Tyrosine phosphorylation of p130(cas) by bombesin, lysophosphatidic acid, phorbol esters, and platelet-derived growth factor. Signaling pathways and formation of a p130(cas)-Crk complex. J. Biol. Chem.272:9363-9370. [DOI] [PubMed] [Google Scholar]
  8. Casamassima, A., and Rozengurt, E. (1998). Insulin-like growth factor I stimulates tyrosine phosphorylation of p130(Cas), focal adhesion kinase, and paxillin. Role of phosphatidylinositol 3′-kinase and formation of a p130(Cas)-Crk complex. J. Biol. Chem.273:26149-26156. [DOI] [PubMed] [Google Scholar]
  9. Chao, M. V. (1992). Growth factor signaling: Where is the specificity? Cell68:995-997. [DOI] [PubMed] [Google Scholar]
  10. Cozzolino, M., Giovannone, B., Serafino, A., Knudsen, K., Levi, A., Alema', S., and Salvatore, A. (2000). Activation of TrkA tyrosine kinase in embryonal carcinoma cells promotes cell compaction, independently of tyrosine phosphorylation of catenins. J. Cell Sci.113:1601-1610. [DOI] [PubMed] [Google Scholar]
  11. Della Rocca, G. J., Maudsley, S., Daaka, Y., Lefkowitz, R. J., and Luttrell, L. M. (1999). Pleiotropic coupling of G protein-coupled receptors to the mitogen-activated protein kinase cascade. Role of focal adhesions and receptor tyrosine kinases. J. Biol. Chem.274:13978-13984. [DOI] [PubMed] [Google Scholar]
  12. Gatti, A. (2003a). Divergence in the upstream signaling of nerve growth factor (NGF) and epidermal growth factor (EGF). NeuroReport14:1031-1035. [DOI] [PubMed] [Google Scholar]
  13. Gatti, A. (2003b). Transient and sustained downregulation of Src family kinases (SFK) in response to epidermal growth factor (EGF) and nerve growth factor (NGF), respectively. Cell. Signal.15:1031-1037. [DOI] [PubMed] [Google Scholar]
  14. Gilmore, A. P., and Romer, L. H. (1996). Inhibition of focal adhesion kinase (FAK) signaling in focal adhesions decreases cell motility and proliferation. Mol. Biol. Cell7:1209-1224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hempstead, B. L., Rabin, S. J., Kaplan, L., Reid, S., Parada, L. F., and Kaplan, D. R. (1992). Overexpression of the trk tyrosine kinase rapidly accelerates nerve growth factor-induced differentiation. Neuron9:883-896. [DOI] [PubMed] [Google Scholar]
  16. Huff, K., End, D., and Guroff, G. (1981). Nerve growth factor-induced alteration in the response of PC12 pheochromocytoma cells to epidermal growth factor. J. Cell Biol.88:189-198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ivankovic-Dikic, I., Gronroos, E., Blaukat, A., Barth, B.-U., and Dikic, I. (2000). Pyk2 and FAK regulate neurite outgrowth induced by growth factors and integrins. Nature Cell Biol.2:574-581. [DOI] [PubMed] [Google Scholar]
  18. Kaplan, D. R., and Miller, F. D. (2000). Neurotrophin signal transduction in the nervous system. Curr. Opin. Neurobiol.10:381-391. [DOI] [PubMed] [Google Scholar]
  19. Kaplan, D. R., and Miller, F. D. (2003). Axon growth inhibition: Signals from the p75 neurotrophin receptor. Nature Neurosci.16:435-436. [DOI] [PubMed] [Google Scholar]
  20. Knight, J. B., Yamauchi, K., and Pessin, J. E. (1995). Divergent insulin and platelet-derived growth factor regulation of focal adhesion kinase (pp125FAK) tyrosine phosphorylation, and rearrangement of actin stress fibers. J. Biol. Chem.270:10199-10203. [DOI] [PubMed] [Google Scholar]
  21. Kremer, N. E., D'arcangelo, G., Thomas, S. M., DeMarco, M., Brugge, J. S., and Halegoua, S. (1991). Signal transduction by nerve growth factor and fibroblast growth factor in PC12 cells requires a sequence of src and ras actions. J. Cell Biol.115:809-819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lazarovici, P., Oshima, M., Shavit, D., Shibutani, M., Jiang, H., Monshipouri, M., Fink, D., Movsesyan, V., and Guroff, G. (1997). Down-regulation of epidermal growth factor receptors by nerve growth factor in PC12 cells is p140(trk)-, Ras-, and Src-dependent. J. Biol. Chem.272:11026-11034. [DOI] [PubMed] [Google Scholar]
  23. Lu, Z., Jiang, G., Blume-Jensen, P., and Hunter, T. (2001). Epidermal growth factor-induced tumor cell invasion and metastasis initiated by dephosphorylation and downregulation of focal adhesion kinase. Mol. Cell. Biol.21:4016-4031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maher, P. A. (1988). Nerve growth factor induces protein-tyrosine phosphorylation. Proc. Natl. Acad. Sci. U.S.A.85:6788-6791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maher, P. A., Pasquale, E. B., Wang, J. Y. J., and Singer, S. J. (1985). Phosphotyrosine-containing proteins are concentrated in focal adhesions and intercellular junctions in normal cells. Proc. Natl. Acad. Sci. U.S.A.82:6576-6580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marshall, C. J. (1995). Specificity of receptor tyrosine kinase signaling: Transient versus sustained extracellular signal-regulated kinase activation. Cell80:179-185. [DOI] [PubMed] [Google Scholar]
  27. Nishiki, T., Narumiya, S., Morii, N., Yamamoto, M., Fujiwara, M., Kamata, Y., Sakaguchi, G., and Kozaki, S. (1990). ADP-ribosylation of the rho/rac proteins induces growth inhibition, neurite outgrowth and acetylcholine esterase in cultured PC12 cells. Biochem. Biophys. Res. Commun.167:265-272. [DOI] [PubMed] [Google Scholar]
  28. Park, S.-Y., Avraham, H., and Avraham, S. (2000). Characterization of the tyrosine kinases RAFTK/Pyk2 and FAK in nerve growth factor-induced neuronal differentiation. J. Biol. Chem.275:19768-19777. [DOI] [PubMed] [Google Scholar]
  29. Parsons, J. T., Martin, K. H., Slack, J. K., Taylor, J. M., and Weed, S. A. (2000). Focal adhesion kinase: A regulator of focal adhesion dynamics and cell movement. Oncogene19:5606-5613. [DOI] [PubMed] [Google Scholar]
  30. Renshaw, M. W., Price, L. S., and Schwartz, A. (1999). Focal adhesion kinase mediates the integrin signaling requirement for growth factor activation of MAP kinase. J. Cell Biol.147:611-618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rodriguez-Fernandez, J. L., and Rozengurt, E. (1996). Bombesin, bradykinin, vasopressin, and phorbol esters rapidly and transiently activate Src family tyrosine kinases in Swiss 3T3 cells. Dissociation from tyrosine phosphorylation of p125 focal adhesion kinase. J. Biol. Chem.271:27895-27901. [DOI] [PubMed] [Google Scholar]
  32. Salazar, E. P., and Rozengurt, E. (2001). Src family kinases are required for integrin-mediated but not for G protein-coupled receptor stimulation of focal adhesion kinase autophosphorylation at Tyr-397. J. Biol. Chem.276:17788-17795. [DOI] [PubMed] [Google Scholar]
  33. Schlessinger, J. (2000). Cell signaling by receptor tyrosine kinases. Cell103:211-225. [DOI] [PubMed] [Google Scholar]
  34. Schubert, D., and Whitlock, C. (1977). Alteration of cellular adhesion by nerve growth factor. Proc. Natl. Acad. Sci. U.S.A.74:4055-4058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thomas, S. M., and Brugge, J. S. (1997). Cellular functions regulated by Src family kinases. Annu. Rev. Cell Dev. Biol.13:513-609. [DOI] [PubMed] [Google Scholar]
  36. Wells, A. (2000). Tumor invasion: Role of growth factor-induced cell motility. Adv. Cancer Res.78:31-101. [DOI] [PubMed] [Google Scholar]
  37. York, R. D., Yao, H., Dillon, T., Ellig, C. L., Eckert, S. P., McCleskey, E. W., and Stork, P. J. S. (1998). Rap1 mediates sustained MAP kinase activation induced by nerve growth factor. Nature392:622-626. [DOI] [PubMed] [Google Scholar]

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