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Focal adhesion kinase controls actin assembly via a FERM-mediated interaction with the Arp2/3 complex

Nature Cell Biology volume 9, pages 1046–1056 (2007)Cite this article

Abstract

Networks of actin filaments, controlled by the Arp2/3 complex, drive membrane protrusion during cell migration. How integrins signal to the Arp2/3 complex is not well understood. Here, we show that focal adhesion kinase (FAK) and the Arp2/3 complex associate and colocalize at transient structures formed early after adhesion. Nascent lamellipodia, which originate at these structures, do not form in FAK-deficient cells, or in cells in which FAK mutants cannot be autophosphorylated after integrin engagement. The FERM domain of FAK binds directly to Arp3 and can enhance Arp2/3-dependent actin polymerization. Critically, Arp2/3 is not bound when FAK is phosphorylated on Tyr 397. Interfering peptides and FERM-domain point mutants show that FAK binding to Arp2/3 controls protrusive lamellipodia formation and cell spreading. This establishes a new function for the FAK FERM domain in forming a phosphorylation-regulated complex with Arp2/3, linking integrin signalling directly with the actin polymerization machinery.

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Figure 1: Formation of early spreading adhesions requires FAK and FAK Tyr 397 phosphorylation.
Figure 2: FAK binds to Arp3 when FAK Tyr 397 is not phosphorylated.
Figure 3: Arp3 no longer colocalizes with FAK as lamellipodia form beyond early spreading adhesions.
Figure 4: Arp3 association with the FAK FERM domain is regulated by FAK Tyr 397 phosphorylation.
Figure 5: Identification of Arp2/3 peptide binding sequences within FAK FERM domain.
Figure 6: Introduction of FERM domain Arp2/3 binding peptides reduces cell spreading and stress fibres.
Figure 7: Introduction of a FERM-domain effector mutant reduces lamellipodia production and cell spreading.
Figure 8: FAK FERM domain enhances Arp2/3-mediated actin polymerization.

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Acknowledgements

This work was supported by Cancer Research UK Program Grant and the Medical Research Council (G.E.J.). We would like to thank M. Kirschner for anti-WASP antibody and D. Ilic for FAK-deficient mouse cells.

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Author notes

  1. Gordon W. McLean

    Present address: Current address: Queen's Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh EH16 4SA, UK.,

Authors and Affiliations

  1. The Beatson Institute for Cancer Research, Cancer Research UK Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, G61 1BD, Glasgow, UK

    Bryan Serrels, Alan Serrels, Valerie G. Brunton, Gordon W. McLean, Christopher H. Gray & Margaret C. Frame

  2. Randall Division of Cell & Molecular Biophysics, King's College London, Guy's Campus, London, SE1 1UL, UK

    Mark Holt & Gareth E. Jones

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Contributions

B.S. and A.S. performed the experimental work. M.H. performed FRAP analysis. V.G.B., G.W.M., G.E.J. and M.C.F. provided extensive scientific input. C.H.G. provided structural biology support. M.C.F. carried out the scientific writing.

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Correspondence to Margaret C. Frame.

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Serrels, B., Serrels, A., Brunton, V. et al. Focal adhesion kinase controls actin assembly via a FERM-mediated interaction with the Arp2/3 complex. Nat Cell Biol 9, 1046–1056 (2007). https://doi.org/10.1038/ncb1626

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