Nanoscale architecture of integrin-based cell adhesions

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Abstract

Cell adhesions to the extracellular matrix (ECM) are necessary for morphogenesis, immunity and wound healing1,2. Focal adhesions are multifunctional organelles that mediate cell–ECM adhesion, force transmission, cytoskeletal regulation and signalling1,2,3. Focal adhesions consist of a complex network4 of trans-plasma-membrane integrins and cytoplasmic proteins that form a <200-nm plaque5,6 linking the ECM to the actin cytoskeleton. The complexity of focal adhesion composition and dynamics implicate an intricate molecular machine7,8. However, focal adhesion molecular architecture remains unknown. Here we used three-dimensional super-resolution fluorescence microscopy (interferometric photoactivated localization microscopy)9 to map nanoscale protein organization in focal adhesions. Our results reveal that integrins and actin are vertically separated by a 40-nm focal adhesion core region consisting of multiple protein-specific strata: a membrane-apposed integrin signalling layer containing integrin cytoplasmic tails, focal adhesion kinase and paxillin; an intermediate force-transduction layer containing talin and vinculin; and an uppermost actin-regulatory layer containing zyxin, vasodilator-stimulated phosphoprotein and α-actinin. By localizing amino- and carboxy-terminally tagged talins, we reveal talin’s polarized orientation, indicative of a role in organizing the focal adhesion strata. The composite multilaminar protein architecture provides a molecular blueprint for understanding focal adhesion functions.

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Figure 1: iPALM imaging of a plasma membrane marker, integrin α v and actin.
Figure 2: Protein stratification of the focal adhesion core.
Figure 3: Talin orientation in focal adhesions.
Figure 4: Nanoscale architecture of focal adhesions.

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Acknowledgements

We thank J. Lippincott-Schwartz, G. Patterson and M. Parsons for sharing DNA; S. Xie for help with automation software; K. Jaqaman for MATLAB code; and HHMI Janelia Farm Scientific Computing and NIH Helix systems for computing resources. Funding: Division of Intramural Research, NHLBI (P.K., A.M.P. and C.M.W.); Howard Hughes Medical Institute (G.S. and H.F.H.).

Author information

P.K. and G.S. collected data and performed data analyses. G.S. and H.F.H. designed and built the instrument. A.M.P. performed immunoprecipitation and western blot experiments. E.B.R. and M.W.D. created expression constructs. P.K., C.M.W., G.S., H.F.H., M.W.D. and A.M.P. wrote the manuscript. P.K. and G.S. contributed equally to the study. All authors discussed the results and commented on the manuscript.

Correspondence to Michael W. Davidson or Harald F. Hess or Clare M. Waterman.

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The authors declare no competing financial interests.

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The file contains Supplementary Figures 1-22 with legends, Supplementary Tables 1-4 and, Supplementary Notes 1-6 and additional references. (PDF 4167 kb)

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Kanchanawong, P., Shtengel, G., Pasapera, A. et al. Nanoscale architecture of integrin-based cell adhesions. Nature 468, 580–584 (2010) doi:10.1038/nature09621

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