Abstract
Vinculin is a conserved component and an essential regulator of both cell–cell (cadherin-mediated) and cell–matrix (integrin–talin-mediated focal adhesions) junctions, and it anchors these adhesion complexes to the actin cytoskeleton by binding to talin in integrin complexes or to α-actinin in cadherin junctions1,2,3. In its resting state, vinculin is held in a closed conformation through interactions between its head (Vh) and tail (Vt) domains4,5,6. The binding of vinculin to focal adhesions requires its association with talin. Here we report the crystal structures of human vinculin in its inactive and talin-activated states. Talin binding induces marked conformational changes in Vh, creating a novel helical bundle structure, and this alteration actively displaces Vt from Vh. These results, as well as the ability of α-actinin to also bind to Vh and displace Vt from pre-existing Vh–Vt complexes, support a model whereby Vh functions as a domain that undergoes marked structural changes that allow vinculin to direct cytoskeletal assembly in focal adhesions and adherens junctions. Notably, talin's effects on Vh structure establish helical bundle conversion as a signalling mechanism by which proteins direct cellular responses.
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Acknowledgements
We thank J. Cleveland for many helpful discussions. We also thank V. Morris, K. Brown and C. Kirby for technical assistance; C. Vonrhein for expert advice and help with autoSHARP; C. Ross for maintaining the X-ray and computing facilities; L. Messerle for the tantalum compound; and M. Kastan for critical review of the manuscript. We are grateful to the staff at the Advanced Photon Source, COM-CAT, SBC-CAT and SER-CAT, and at the Advanced Light Source, Lawrence Berkeley Laboratory, 5.0.2, for synchrotron support. This work was supported in part by the Cancer Center Support (CORE) Grant and by the American Lebanese Syrian Associated Charities (ALSAC). P.B. is a Van Vleet Fellow.
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Supplementary information
41586_2004_BFnature02281_MOESM10_ESM.doc
Supplementary Table 2: Vh residues interacting with talin VBS3 residues as seen in the Vh:VBS3 crystal structure. (DOC 31 kb)
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Izard, T., Evans, G., Borgon, R. et al. Vinculin activation by talin through helical bundle conversion. Nature 427, 171–175 (2004). https://doi.org/10.1038/nature02281
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DOI: https://doi.org/10.1038/nature02281
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