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Dissecting the molecular architecture of integrin adhesion sites by cryo-electron tomography

Nature Cell Biology volume 12, pages 909–915 (2010)Cite this article

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Abstract

Focal adhesions are integrin-based multiprotein complexes, several micrometres in diameter, that mechanically link the extracellular matrix with the termini of actin bundles. The molecular diversity of focal adhesions and their role in cell migration and matrix sensing has been extensively studied, but their ultrastructural architecture is still unknown. We present the first three-dimensional structural reconstruction of focal adhesions using cryo-electron tomography. Our analyses reveal that the membrane–cytoskeleton interaction at focal adhesions is mediated through particles located at the cell membrane and attached to actin fibres. The particles have diameters of 25 ± 5 nm, and an average interspacing of approximately 45 nm. Treatment with the Rho-kinase inhibitor Y-27632 induces a rapid decrease in particle diameter, suggesting that they are highly mechanosensitive. Our findings clarify the internal architecture of focal adhesions at molecular resolution, and provide insights into their scaffolding and mechanosensory functions.

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Figure 1: Cryo-electron tomography of integrin-mediated focal adhesions.
Figure 2: Adhesion-related particles at sites of focal adhesions.
Figure 3: Heterogeneity of adhesion-related particles and vinculin localization.
Figure 4: Molecular changes in the adhesion machinery induced by the Rho-kinase inhibitor Y-27632.
Figure 5: Architecture of focal adhesions at nanoscale resolution.

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  • 17 August 2010

    In the version of this letter initially published online, Vera Hirschfeld-Warneken was incorrectly spelled in the author list. This error has been corrected in both the HTML and PDF versions of the letter.

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Acknowledgements

This study was supported by a grant from the German-Israeli Cooperation Project (DIP H.2.2) to O. M., R. F., J. S. and B. G., an NIGMS grant from the National Institutes of Health Cell Migration Consortium (Grant No. U54 GM64346) to B. G. and an ERC Starting Grant to O. M. The authors express gratitude to B. Morgenstern for help in editing the manuscript. B. G. holds the Erwin Neter Professorial Chair in Cell and Tumor Biology.

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Authors and Affiliations

  1. Department of Life Sciences and the National Institute for Biotechnology in the Negev, Ben Gurion University of the Negev, Be'er-Sheva, 84120, Israel

    Israel Patla, Nadav Elad & Ohad Medalia

  2. Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, 76100, Israel

    Tova Volberg & Benjamin Geiger

  3. Department of New Materials and Biosystems, Max-Planck Institute for Metals Research, Heisenbergstrasse, Stuttgart, 70569, Germany

    Vera Hirschfeld-Warneken & Joachim P. Spatz

  4. Department of Biophysical Chemistry, University of Heidelberg, Stuttgart, D-70569, Germany

    Vera Hirschfeld-Warneken

  5. Department of Molecular Medicine, Max-Planck Institute of Biochemistry, Martinsried, D-82152, Germany

    Carsten Grashoff & Reinhard Fässler

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Contributions

I. P., N. E., T. V., C. G. and V. H. W. performed experimental work and analysed the data. B. G., J. S., R. F. and O. M. designed the experiments, analysed the data and prepared the manuscript.

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Correspondence to Ohad Medalia.

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Patla, I., Volberg, T., Elad, N. et al. Dissecting the molecular architecture of integrin adhesion sites by cryo-electron tomography. Nat Cell Biol 12, 909–915 (2010). https://doi.org/10.1038/ncb2095

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