This Article Full Text Full Text (PDF) Supplementary Data Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Brunton, V. G. Articles by Frame, M. C. Search for Related Content PubMed PubMed Citation Articles by Brunton, V. G. Articles by Frame, M. C.

Identification of Src-Specific Phosphorylation Site on Focal Adhesion Kinase: Dissection of the Role of Src SH2 and Catalytic Functions and Their Consequences for Tumor Cell Behavior

¹ Beatson Institute for Cancer Research, Cancer Research UK Beatson Laboratories, Glasgow, United Kingdom and ² ARIAD Pharmaceuticals, Cambridge, Massachusetts

Requests for reprints: Valerie G. Brunton, Beatson Institute for Cancer Research, Cancer Research UK Beatson Laboratories, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK. Phone: 44-141-330-3956; Fax: 44-141-942-6521; E-mail: v.brunton{at}beatson.gla.ac.uk

Src tyrosine kinase expression and activity are elevated during colon cancer progression. How this contributes to the malignant phenotype is not fully understood. We show that in KM12C colon carcinoma cells, expression of kinase-deficient Src proteins (SrcMF and Src251) does not alter cell growth. Src kinase activity is required for turnover of cell-matrix adhesions and, in particular, the Src-dependent phosphorylation of focal adhesion kinase (FAK) is required for their disassembly. Surprisingly, we found that expression of SrcMF or Src251 resulted in increased tyrosine phosphorylation of FAK on Tyr⁴⁰⁷, Tyr⁵⁷⁶, Tyr⁵⁷⁷, and Tyr⁸⁶¹, which are considered to be Src kinase substrates. This Src kinase–independent phosphorylation of FAK required an intact Src SH2 domain that mediates association of Src and FAK at peripheral adhesions. Use of a novel highly potent and selective Src kinase inhibitor AP23464 combined with experiments in Src/Fyn/Yes–deficient fibroblasts showed that increased phosphorylation of FAK in cells expressing SrcMF did not require Src-like kinases. However, specific phosphorylation on Tyr⁹²⁵ of FAK was not evident in SrcMF- or Src251-expressing cells, and lack of Src kinase–dependent phosphorylation on this site was associated with impaired adhesion turnover. Our data show that Src kinase activity is required for adhesion turnover associated with cell migration in cancer cells and that, in addition to the catalytic activity, Src also acts as an adaptor to recruit other kinases that can phosphorylate key substrates including FAK. These studies have implications for tumor progression with respect to the use of Src kinase inhibitors.

Key Words: Src kinase • focal adhesion kinase • colon cancer • cell migration

This article has been cited by other articles:

				K. Ren, H. Jin, C. Bian, H. He, X. Liu, S. Zhang, Y. Wang, and R.-g. Shao MR-1 Modulates Proliferation and Migration of Human Hepatoma HepG2 Cells through Myosin Light Chains-2 (MLC2)/Focal Adhesion Kinase (FAK)/Akt Signaling Pathway J. Biol. Chem., December 19, 2008; 283(51): 35598 - 35605. [Abstract] [Full Text] [PDF]

				A. Vultur, R. Buettner, C. Kowolik, W. Liang, D. Smith, F. Boschelli, and R. Jove SKI-606 (bosutinib), a novel Src kinase inhibitor, suppresses migration and invasion of human breast cancer cells Mol. Cancer Ther., May 1, 2008; 7(5): 1185 - 1194. [Abstract] [Full Text] [PDF]

				A. Spreafico, S. Schenone, T. Serchi, M. Orlandini, A. Angelucci, D. Magrini, G. Bernardini, G. Collodel, A. Di Stefano, C. Tintori, et al. Antiproliferative and proapoptotic activities of new pyrazolo[3,4-d]pyrimidine derivative Src kinase inhibitors in human osteosarcoma cells FASEB J, May 1, 2008; 22(5): 1560 - 1571. [Abstract] [Full Text] [PDF]

				T. Hayashida, M.-H. Wu, A. Pierce, A.-C. Poncelet, J. Varga, and H. W. Schnaper MAP-kinase activity necessary for TGFbeta1-stimulated mesangial cell type I collagen expression requires adhesion-dependent phosphorylation of FAK tyrosine 397 J. Cell Sci., December 1, 2007; 120(23): 4230 - 4240. [Abstract] [Full Text] [PDF]

				Y. Lim, H. Park, J. Jeon, I. Han, J. Kim, E.-H. Jho, and E.-S. Oh Focal Adhesion Kinase Is Negatively Regulated by Phosphorylation at Tyrosine 407 J. Biol. Chem., April 6, 2007; 282(14): 10398 - 10404. [Abstract] [Full Text] [PDF]

				A. Di Florio, G. Capurso, M. Milione, F. Panzuto, R. Geremia, G. D. Fave, and C. Sette Src family kinase activity regulates adhesion, spreading and migration of pancreatic endocrine tumour cells Endocr. Relat. Cancer, March 1, 2007; 14(1): 111 - 124. [Abstract] [Full Text] [PDF]

				H. Achiwa and J. S. Lazo PRL-1 Tyrosine Phosphatase Regulates c-Src Levels, Adherence, and Invasion in Human Lung Cancer Cells Cancer Res., January 15, 2007; 67(2): 643 - 650. [Abstract] [Full Text] [PDF]

				A. Serrels, I. R.J. Macpherson, T.R. J. Evans, F. Y. Lee, E. A. Clark, O. J. Sansom, G. H. Ashton, M. C. Frame, and V. G. Brunton Identification of potential biomarkers for measuring inhibition of Src kinase activity in colon cancer cells following treatment with dasatinib Mol. Cancer Ther., December 1, 2006; 5(12): 3014 - 3022. [Abstract] [Full Text] [PDF]

				Y.-P. Chong, A. S. Chan, K.-C. Chan, N. A. Williamson, E. C. Lerner, T. E. Smithgall, J. D. Bjorge, D. J. Fujita, A. W. Purcell, G. Scholz, et al. C-terminal Src Kinase-homologous Kinase (CHK), a Unique Inhibitor Inactivating Multiple Active Conformations of Src Family Tyrosine Kinases J. Biol. Chem., November 3, 2006; 281(44): 32988 - 32999. [Abstract] [Full Text] [PDF]

				A. Gampel, L. Moss, M. C. Jones, V. Brunton, J. C. Norman, and H. Mellor VEGF regulates the mobilization of VEGFR2/KDR from an intracellular endothelial storage compartment Blood, October 15, 2006; 108(8): 2624 - 2631. [Abstract] [Full Text] [PDF]

				L. Gonzalez, M. T. Agullo-Ortuno, J. M. Garcia-Martinez, A. Calcabrini, C. Gamallo, J. Palacios, A. Aranda, and J. Martin-Perez Role of c-Src in Human MCF7 Breast Cancer Cell Tumorigenesis J. Biol. Chem., July 28, 2006; 281(30): 20851 - 20864. [Abstract] [Full Text] [PDF]

				S. Meierjohann, E. Wende, A. Kraiss, C. Wellbrock, and M. Schartl The oncogenic epidermal growth factor receptor variant xiphophorus melanoma receptor kinase induces motility in melanocytes by modulation of focal adhesions. Cancer Res., March 15, 2006; 66(6): 3145 - 3152. [Abstract] [Full Text] [PDF]

				J. M. Summy and G. E. Gallick Treatment for Advanced Tumors: Src Reclaims Center Stage Clin. Cancer Res., March 1, 2006; 12(5): 1398 - 1401. [Full Text] [PDF]

				S. J. Cohen, R. B. Cohen, and N. J. Meropol Targeting Signal Transduction Pathways in Colorectal Cancer--More Than Skin Deep J. Clin. Oncol., August 10, 2005; 23(23): 5374 - 5385. [Abstract] [Full Text] [PDF]