FAK dimerization controls its kinase-dependent functions at focal adhesions

Karen Brami-Cherrier; Nicolas Gervasi; Diana A. Arsenieva; Katarzyna Walkiewicz; Marie Claude Boutterin; Álvaro Darío Ortega; Paul G. Leonard; Bastien Seantier; Laïla Gasmi; Tahar Bouceba; Gress Kadaré; Jean Antoine Girault -; Stefan T. Arold

doi:10.1002/embj.201386399

FAK dimerization controls its kinase-dependent functions at focal adhesions

Karen Brami-Cherrier, Nicolas Gervasi, Diana A. Arsenieva, Katarzyna Walkiewicz, Marie Claude Boutterin, Álvaro Darío Ortega, Paul G. Leonard, Bastien Seantier, Laïla Gasmi, Tahar Bouceba, Gress Kadaré, Jean Antoine Girault -, Stefan T. Arold

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104 Scopus citations

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.

Original language	English (US)
Pages (from-to)	356-370
Number of pages	15
Journal	The EMBO Journal
Volume	33
Issue number	4
DOIs	https://doi.org/10.1002/embj.201386399
State	Published - Jan 30 2014

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
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.

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
Molecular Biology
General Immunology and Microbiology

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10.1002/embj.201386399

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@article{7d7d6630157f477dab9e94ca67874140,

title = "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. {\textcopyright} 2014 The Authors.",

author = "Karen Brami-Cherrier and Nicolas Gervasi and Arsenieva, {Diana A.} and Katarzyna Walkiewicz and Boutterin, {Marie Claude} and Ortega, {{\'A}lvaro Dar{\'i}o} and Leonard, {Paul G.} and Bastien Seantier and La{\"i}la Gasmi and Tahar Bouceba and Gress Kadar{\'e} and {Girault -}, {Jean Antoine} and Arold, {Stefan T.}",

note = "KAUST Repository Item: Exported on 2020-10-01 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.",

year = "2014",

month = jan,

day = "30",

doi = "10.1002/embj.201386399",

language = "English (US)",

volume = "33",

pages = "356--370",

journal = "The EMBO Journal",

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TY - JOUR

T1 - FAK dimerization controls its kinase-dependent functions at focal adhesions

AU - Brami-Cherrier, Karen

AU - Gervasi, Nicolas

AU - Arsenieva, Diana A.

AU - Walkiewicz, Katarzyna

AU - Boutterin, Marie Claude

AU - Ortega, Álvaro Darío

AU - Leonard, Paul G.

AU - Seantier, Bastien

AU - Gasmi, Laïla

AU - Bouceba, Tahar

AU - Kadaré, Gress

AU - Girault -, Jean Antoine

AU - Arold, Stefan T.

N1 - KAUST Repository Item: Exported on 2020-10-01 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.

PY - 2014/1/30

Y1 - 2014/1/30

N2 - 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.

AB - 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.

UR - http://hdl.handle.net/10754/563361

UR - http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3989642

UR - http://www.scopus.com/inward/record.url?scp=84897862926&partnerID=8YFLogxK

U2 - 10.1002/embj.201386399

DO - 10.1002/embj.201386399

M3 - Article

C2 - 24480479

SN - 0261-4189

VL - 33

SP - 356

EP - 370

JO - The EMBO Journal

JF - The EMBO Journal

IS - 4

ER -

FAK dimerization controls its kinase-dependent functions at focal adhesions

Abstract

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