FAK dimerization controls its kinase-dependent functions at focal adhesions

Abstract
Focal adhesion kinase (FAK) controls adhesion-dependent cell motility, survival, and proliferation. FAK has kinase-dependent and kinase-independent functions, both of which play major roles in embryogenesis and tumor invasiveness. The precise mechanisms of FAK activation are not known. Using x-ray crystallography, small angle x-ray scattering, and biochemical and functional analyses, we show that the key step for activation of FAK's kinase-dependent functions-autophosphorylation of tyrosine-397-requires site-specific dimerization of FAK. The dimers form via the association of the N-terminal FERM domain of FAK and are stabilized by an interaction between FERM and the C-terminal FAT domain. FAT binds to a basic motif on FERM that regulates co-activation and nuclear localization. FAK dimerization requires local enrichment, which occurs specifically at focal adhesions. Paxillin plays a dual role, by recruiting FAK to focal adhesions and by reinforcing the FAT:FERM interaction. Our results provide a structural and mechanistic framework to explain how FAK combines multiple stimuli into a site-specific function. The dimer interfaces we describe are promising targets for blocking FAK activation. © 2014 The Authors.

Citation
Brami-Cherrier, K., Gervasi, N., Arsenieva, D., Walkiewicz, K., Boutterin, M.-C., Ortega, A., … Arold, S. T. (2014). FAK dimerization controls its kinase-dependent functions at focal adhesions. The EMBO Journal, 33(4), 356–370. doi:10.1002/embj.201386399

Acknowledgements
We thank S. Lachkar for help with SEC, K. Muller and V. Unkefer for editorial assistance, J.E. Ladbury for access to the MicroCal iTC200, L. Ponchon for help with DLS, D. Svergun and his colleagues for assistance with SAXS data recording and analysis at the x33 beamline at the European Molecular Biology Laboratory/Deutsches Elektronen-Synchrotron, G. Meigs for help with X-ray crystallography data recording at the Advanced Light Source beamline bl.8.3.1., Berkeley, CA, D. Ilic for the gift of Ptk2-/- fibroblasts, and A. Sobel and R.M. Mege for critical reading of the manuscript. We acknowledge support with data recording at the European Synchrotron Radiation Facility beamline ID14 and from the European Community Research Infrastructure Action under the Sixth Framework Program (RII3/CT/2004/5060008) for access to the European Molecular Biology Laboratory/Deutsches Elektronen-Synchrotron. This work was supported by Agence Nationale de la Recherche (ANR-05-2_42589), Association pour la Recherche sur le Cancer (ARC, A05/3/3138), Fondation pour la Recherche Medicale, European Research Council, Inserm, the University Cancer Foundation via the Institutional Research Grant program at the University of Texas MD Anderson Cancer Center, by NIH/NCI grant R03 CA169969-01, and, in part, by the National Institutes of Health through MD Anderson's Cancer Center Support Grant (CA016672). KBC was recipient of fellowships from ARC and Region Ile de France (NeRF). J.A. Girault's group is affiliated with the Ecole des Neurosciences de Paris-Ile-de-France and the Bio-Psy Laboratory of Excellence.

Publisher
Wiley

Journal
The EMBO Journal

DOI
10.1002/embj.201386399

PubMed ID
24480479

PubMed Central ID
PMC3989642

Additional Links
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3989642

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