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FERM domain interaction with myosin negatively regulates FAK in cardiomyocyte hypertrophy

Nature Chemical Biology volume 8, pages 102–110 (2012)Cite this article

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Abstract

Focal adhesion kinase (FAK) regulates cellular processes that affect several aspects of development and disease. The FAK N-terminal FERM (4.1 protein–ezrin-radixin-moesin homology) domain, a compact clover-leaf structure, binds partner proteins and mediates intramolecular regulatory interactions. Combined chemical cross-linking coupled to MS, small-angle X-ray scattering, computational docking and mutational analyses showed that the FAK FERM domain has a molecular cleft (∼998 Å2) that interacts with sarcomeric myosin, resulting in FAK inhibition. Accordingly, mutations in a unique short amino acid sequence of the FERM myosin cleft, FP-1, impaired the interaction with myosin and enhanced FAK activity in cardiomyocytes. An FP-1 decoy peptide selectively inhibited myosin interaction and increased FAK activity, promoting cardiomyocyte hypertrophy through activation of the AKT–mammalian target of rapamycin pathway. Our findings uncover an inhibitory interaction between the FAK FERM domain and sarcomeric myosin that presents potential opportunities to modulate the cardiac hypertrophic response through changes in FAK activity.

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Figure 1: Interaction and regulation of FAK by cardiac myosin in vitro.
Figure 2: FAK-myosin interacting surface determined by CXMS and SAXS.
Figure 3: Mutagenesis of the FERM-myosin complex interface.
Figure 4: FP-1 interacts with sarcomeric myosin and negatively regulates the FAK-myosin interaction in vitro.
Figure 5: FAK interacts with native myosin in cardiomyocytes.
Figure 6: Stretch or treatment with FP-1-TAT inhibits FAK-myosin interaction and induces FAK phosphorylation in NRVMs.
Figure 7: Activation of FAK by treatment with FP-1-TAT induces hypertrophy in NRVMs via the AKT-mTOR signaling pathway.

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Acknowledgements

This work was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP; Grants 2006/54878-3, 2007/55930-1, 2007/59442-1, 2008/53519-5, 2008/57805-2, 2010/02628-9) and Conselho Nacional de Pesquisa (CNPq: Grants 304366/2009-9, 475158/2010-5, 573672/2008-3, 559698/2009-7).

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Authors and Affiliations

  1. Department of Internal Medicine, School of Medicine, University of Campinas, Campinas, São Paulo, Brazil.,

    Aline M Santos, Alisson C Cardoso, Michelle B M Pereira, Talita M Marin & Kleber G Franchini

  2. Brazilian National Laboratory for Biosciences, Brazilian Association for Synchrotron Light Technology, Campinas, São Paulo, Brazil.,

    Aline M Santos, Alisson C Cardoso, Carolina F M Z Clemente, Júlio C Silva, Michelle B M Pereira, Talita M Marin, Paulo S L Oliveira, Ana Carolina M Figueira, Saulo H P Oliveira, Íris L Torriani, José Xavier Neto & Kleber G Franchini

  3. Department of Biochemistry, Chemistry Institute, University of São Paulo, São Paulo, Brazil

    Deborah Schechtman

  4. Chemistry Institute, University of Campinas, Campinas, São Paulo, Brazil

    Mariana Fioramonte & Fábio C Gozzo

  5. Gleb Wataghin Physics Institute, University of Campinas, Campinas, São Paulo, Brazil.,

    Íris L Torriani

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  1. Aline M Santos
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Contributions

A.M.S., D.S. and K.G.F. conceived the research and designed experiments. A.M.S., D.S., A.C.C., M.B.M.P. and T.M.M. conducted experiments. A.M.S. and C.F.M.Z.C. designed and performed mutational experiments. A.M.S. and A.C.M.F. performed affinity experiments. A.M.S., J.C.S. and I.L.T. performed and analyzed SAXS experiments. A.M.S., M.F. and F.C.G. performed crosslinking and MS experiments and analysis. A.M.S., P.S.L.O. and S.H.P.O. performed molecular modeling and docking. A.M.S., D.S., J.X.N. and K.G.F. wrote the paper.

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Correspondence to Kleber G Franchini.

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Santos, A., Schechtman, D., Cardoso, A. et al. FERM domain interaction with myosin negatively regulates FAK in cardiomyocyte hypertrophy. Nat Chem Biol 8, 102–110 (2012). https://doi.org/10.1038/nchembio.717

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