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Talin phosphorylation by Cdk5 regulates Smurf1-mediated talin head ubiquitylation and cell migration

Nature Cell Biology volume 11, pages 624630 (2009) | Download Citation

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Abstract

Cell migration is a dynamic process that requires temporal and spatial regulation of integrin activation and focal adhesion assembly/disassembly1. Talin, an actin and β-integrin tail-binding protein, is essential for integrin activation and focal adhesion formation2,3. Calpain-mediated cleavage of talin has a key role in focal adhesion turnover3; however, the talin head domain, one of the two cleavage products, stimulates integrin activation, localizes to focal adhesions and maintains cell edge protrusions2,4,5, suggesting that other steps, downstream of talin proteolysis, are required for focal adhesion disassembly. Here we show that talin head binds Smurf1, an E3 ubiquitin ligase involved in cell polarity and migration6,7, more tightly than full-length talin does and that this interaction leads to talin head ubiquitylation and degradation. We found that talin head is a substrate for Cdk5, a cyclin-dependent protein kinase that is essential for cell migration, synaptic transmission and cancer metastasis8,9,10,11. Cdk5 phosphorylated talin head at Ser 425, inhibiting its binding to Smurf1, thus preventing talin head ubiquitylation and degradation. Expression of the mutant talS425A, which resists Cdk5 phosphorylation thereby increasing its susceptibility to Smurf1-mediated ubiqitylation, resulted in extensive focal adhesion turnover and inhibited cell migration. Thus, talin head produced by calpain-induced cleavage of talin is degraded through Smurf1-mediated ubiquitylation; moreover, phosphorylation by Cdk5 regulates the binding of Smurf1 to talin head, controlling talin head turnover, adhesion stability and ultimately, cell migration.

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Acknowledgements

We thank K. Burridge for critical reading of this manuscript, M. Schaller for critical reading of this manuscript and supporting of phosphorylation assays, A. Huttenlocher for the pEGFP–talin plasmid, L.-H. Tsai for the Cdk5 and p35 plasmids, D. Cyr for the HA-ubiquitin plasmid, E1 and Ubc5α, and M. Kerber for assistance with TIRF imaging. Supported by National Institutes of Health grants to M.H.G., a Cell Migration Consortium grant (NIH GM64346) to M.H.G. and K.J. and a Ruth L. Kirschstein National Research Service Award (1F32 HL08321) to C.H.

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Affiliations

  1. Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599, USA.

    • Cai Huang
    • , Zenon Rajfur
    • , Zaozao Chen
    •  & Ken Jacobson
  2. Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599, USA.

    • Ken Jacobson
  3. Department of Medicine, University of California San Diego, La Jolla, CA 92093, USA.

    • Cai Huang
    • , Nima Yousefi
    •  & Mark H. Ginsberg

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Contributions

C.H. carried out most of the experiments; Z.R. asissted with TIRF imaging, N.Y. purified talin and fragments and Z.C. made Smurf1 mutants and analysed binding to talin; M.H.G., K.J. and C.H. guided the research and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Ken Jacobson or Mark H. Ginsberg.

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https://doi.org/10.1038/ncb1868

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