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Vinculin activation by talin through helical bundle conversion

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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Figure 1: The closed conformation of the human vinculin tail.
Figure 2: Crystal structure of inactive human Vh (pink, residues 1–258; left panel) compared to α-catenin (light yellow, residues 57–261) bound to β-catenin (green, residues 118–149 (PDB 1dow); right panel).
Figure 3: Structure of inactive human vinculin.
Figure 4: Structure of Vh when activated by talin.
Figure 5: Vinculin activation in focal adhesions and adherens junctions.

References

  1. Zamir, E. & Geiger, B. Molecular complexity and dynamics of cell-matrix adhesions. J. Cell Sci. 114, 3583–3590 (2001)

    CAS  PubMed  Google Scholar 

  2. Pokutta, S. & Weis, W. I. The cytoplasmic face of cell contact sites. Curr. Opin. Struct. Biol. 12, 255–262 (2002)

    CAS  Article  PubMed  Google Scholar 

  3. Xu, W., Baribault, H. & Adamson, E. D. Vinculin knockout results in heart and brain defects during embryonic development. Development 125, 327–337 (1998)

    CAS  PubMed  Google Scholar 

  4. Winkler, J., Lunsdorf, H. & Jockusch, B. M. The ultrastructure of chicken gizzard vinculin as visualized by high-resolution electron microscopy. J. Struct. Biol. 116, 270–277 (1996)

    CAS  Article  PubMed  Google Scholar 

  5. Gilmore, A. P. & Burridge, K. Regulation of vinculin binding to talin and actin by phosphatidyl-inositol-4–5-bisphosphate. Nature 381, 531–535 (1996)

    ADS  CAS  Article  PubMed  Google Scholar 

  6. Miller, G. J., Dunn, S. D. & Ball, E. H. Interaction of the N- and C-terminal domains of vinculin. Characterization and mapping studies. J. Biol. Chem. 276, 11729–11734 (2001)

    CAS  Article  PubMed  Google Scholar 

  7. McGregor, A., Blanchard, A. D., Rowe, A. J. & Critchley, D. R. Identification of the vinculin-binding site in the cytoskeletal protein α-actinin. Biochem. J. 301, 225–233 (1994)

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. Johnson, R. P. & Craig, S. W. F-actin binding site masked by the intramolecular association of vinculin head and tail domains. Nature 373, 261–264 (1995)

    ADS  CAS  Article  PubMed  Google Scholar 

  9. Wood, C. K., Turner, C. E., Jackson, P. & Critchley, D. R. Characterization of the paxillin-binding site and the C-terminal focal adhesion targeting sequence in vinculin. J. Cell Sci. 107, 709–717 (1994)

    CAS  PubMed  Google Scholar 

  10. Bakolitsa, C., de Pereda, J. M., Bagshaw, C. R., Critchley, D. R. & Liddington, R. C. Crystal structure of the vinculin tail suggests a pathway for activation. Cell 99, 603–613 (1999)

    CAS  Article  PubMed  Google Scholar 

  11. Bass, M. D., Smith, B. J., Prigent, S. A. & Critchley, D. R. Talin contains three similar vinculin-binding sites predicted to form an amphipathic helix. Biochem. J. 341, 257–263 (1999)

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. Ling, K., Doughman, R. L., Firestone, A. J., Bunce, M. W. & Anderson, R. A. Type I gamma phosphatidylinositol phosphate kinase targets and regulates focal adhesions. Nature 420, 89–93 (2002)

    ADS  CAS  Article  PubMed  Google Scholar 

  13. Di Paolo, G. et al. Recruitment and regulation of phosphatidylinositol phosphate kinase type 1 gamma by the FERM domain of talin. Nature 420, 85–89 (2002)

    ADS  CAS  Article  PubMed  Google Scholar 

  14. Pokutta, S. & Weiss, W. I. Structure of the dimerization and β-catenin-binding region of α-catenin. Mol. Cell 5, 533–543 (2000)

    CAS  Article  PubMed  Google Scholar 

  15. Yang, J., Dokurno, P., Tonks, N. K. & Barford, D. Crystal structure of the M-fragment of α-catenin: implications for modulation of cell adhesion. EMBO J. 20, 3645–3656 (2001)

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. Pokutta, S., Drees, F., Takai, Y., Nelson, W. J. & Weis, W. I. Biochemical and structural definition of the l-afadin- and actin-binding sites of α-catenin. J. Biol. Chem. 277, 18868–18874 (2002)

    CAS  Article  PubMed  Google Scholar 

  17. Weiss, E. E., Kroemker, M., Rudiger, A.-H., Jockusch, B. M. & Rudiger, M. Vinculin is part of the cadherin-catenin junctional complex: complex formation between α-catenin and vinculin. J. Cell Biol. 131, 755–764 (1997)

    Google Scholar 

  18. Watabe-Uchida, M. et al. α-catenin-vinculin interaction functions to organize the apical junctional complex in epithelial cells. J. Cell Biol. 142, 847–857 (1998)

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. Johnson, R. P. & Craig, S. W. An intramolecular association between the head and tail domains of vinculin modulates talin binding. J. Biol. Chem. 269, 12611–12619 (1994)

    CAS  PubMed  Google Scholar 

  20. Bass, M. D. et al. Further characterization of the interaction between the cytoskeletal proteins talin and vinculin. Biochem. J. 362, 761–768 (2002)

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  21. Lawrence, M. C. & Colman, P. M. Shape complementarity at protein/protein interfaces. J. Mol. Biol. 234, 946–950 (1993)

    CAS  Article  PubMed  Google Scholar 

  22. Johnson, R. P. & Craig, S. W. An intramolecular association between the head and tail domains of vinculin modulates talin binding. J. Biol. Chem. 269, 12611–12619 (1994)

    CAS  PubMed  Google Scholar 

  23. Hayashi, I., Vuori, K. & Liddington, R. C. The focal adhesion targeting (FAT) region of focal adhesion kinase is a four-helical bundle that binds paxillin. Nature Struct. Biol. 9, 101–106 (2002)

    CAS  Article  PubMed  Google Scholar 

  24. Hoellerer, M. K. et al. Molecular recognition of paxillin LD motifs by focal adhesion targeting domain. Structure 11, 1207–1217 (2003)

    CAS  Article  PubMed  Google Scholar 

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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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Correspondence to Tina Izard.

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Supplementary information

Supplementary Figure S1 (JPG 164 kb)

Supplementary Figure S2a (JPG 84 kb)

Supplementary Figure S2b (JPG 160 kb)

Supplementary Figure S3 (JPG 38 kb)

Supplementary Figure S4a (JPG 52 kb)

Supplementary Figure S4b (JPG 25 kb)

Supplementary Figure S4c (JPG 44 kb)

Supplementary Figure Legends (DOC 22 kb)

Supplementary Table 1: Vh residues interacting with Vt residues as seen in the Vh:Vt crystal structure. (DOC 30 kb)

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)

Supplementary Table 3: Vh:Vt crystallographic data statistics. (DOC 50 kb)

Supplementary Table 4: Vh:VBS3 crystallographic data statistics. (DOC 50 kb)

Supplementary Table 5: Vh:Vt phasing statistics. (DOC 32 kb)

Supplementary Table 6: Vh:Vt crystallographic refinement statistics against native data. (DOC 24 kb)

Supplementary Table 7: Structure determination, refinement and phasing statistics of the Vh:VBS3 complex. (DOC 28 kb)

Supplementary Table 8: Vh:VBS3 crystallographic refinement statistics against SeMet and native data. (DOC 25 kb)

Supplementary Table References (DOC 21 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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