Article

Fibronectin-bound α5β1 integrins sense load and signal to reinforce adhesion in less than a second

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Abstract

Integrin-mediated mechanosensing of the extracellular environment allows cells to control adhesion and signalling. Whether cells sense and respond to force immediately upon ligand-binding is unknown. Here, we report that during adhesion initiation, fibroblasts respond to mechanical load by strengthening integrin-mediated adhesion to fibronectin (FN) in a biphasic manner. In the first phase, which depends on talin and kindlin as well as on the actin nucleators Arp2/3 and mDia, FN-engaged α5β1 integrins activate focal adhesion kinase (FAK) and c-Src in less than 0.5 s to steeply strengthen α5β1- and αV-class integrin-mediated adhesion. When the mechanical load exceeds a certain threshold, fibroblasts decrease adhesion and initiate the second phase, which is characterized by less steep adhesion strengthening. This unique, biphasic cellular adhesion response is mediated by α5β1 integrins, which form catch bonds with FN and signal to FN-binding integrins to reinforce cell adhesion much before visible adhesion clusters are formed.

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Acknowledgements

We thank S. Weiser and J. Thoma for fibronectin fragment cloning, expression and purification, the Single Cell Facility (SCF) of the Department of Biosystems Science and Engineering of the ETH Zürich for help with flow cytometry, N. Beerenwinkel for help with statistical questions and J. Helenius and J. Polleux for critical discussions. This work was supported by the European Research Council (Grant Agreement no. 322652), Deutsche Forschungsgemeinschaft (SFB865-B3), and Swiss National Science Foundation (Grant 31003A_138063).

Author information

Affiliations

  1. Eidgenössische Technische Hochschule (ETH) Zürich, Department of Biosystems Science and Engineering, 4058 Basel, Switzerland

    • Nico Strohmeyer
    • , Mitasha Bharadwaj
    •  & Daniel J. Müller
  2. Department of Biochemistry and Molecular Biology, Estructura de Reserca Interdisciplinar en Biotechnologia i Biomedicina, Universitat de València, 46100 Burjassot, Spain

    • Mercedes Costell
  3. Max Planck Institute of Biochemistry, Department of Molecular Medicine, 82152 Martinsried, Germany

    • Reinhard Fässler

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Contributions

N.S., M.B., R.F. and D.J.M. designed the experiments and wrote the paper. N.S. performed and analysed most experiments. M.B. performed initial experiments. R.F. and M.C. provided reagents and/or analytical tools. All authors discussed the experiments, read and approved the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Daniel J. Müller.

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