Abstract
ALTHOUGH vinculin is present at all sites of F-actin attachment to plasma membranes1 and is required for linkage of myofibrils to sarcolemma2, it is unclear how it promotes attachment of actin to membranes. Because biochemical evidence for a direct interaction of vinculin with F-actin is controversial3–9, current models of actin-membrane linkages depict only an indirect role for vinculin, as a tether for α-actinin10. We demonstrate here that an intramolecular association between the 95K head and 30K tail domains of vinculin11 masks an F-actin binding site present in the carboxy-terminal tail domain. Cosedimentation and crosslinking assays, and direct visualization by transmission electron microscopy, reveal an interaction between F-actin and a bacterially expressed fusion protein containing amino acids 811–1066 of vinculin, and between F-actin and a proteolytic fragment of vinculin containing amino acids 858-1066. Vinculin itself neither cosediments with nor crosslinks F-actin. The amino-terminal 95K head fragment of vinculin, but not intact vinculin, inhibits both cosedimentation and crosslinking. We propose that assembly of vinculin into an adherens junction involves disruption of the head-tail interaction, revealing a site that mediates microfilament attachment.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Geiger, B., Tokuyasu, K. T., Dutton, A. H. & Singer, S. J. Proc. natn. Acad. Sci. U.S.A. 77, 4127–4131 (1980).
Barstead, R. J. & Waterston, R. H. J. Cell Biol. 114, 715–724 (1991).
Jockusch, B. M. & Isenberg, G. Proc. natn. Acad. Sci. U.S.A. 78, 3005–3009 (1981).
Wilkins, J. A. & Lin, S. Cell 28, 83–90 (1982).
Evans, R. R., Robson, R. M. & Stromer, M. H. J. biol. Chem. 259, 3916–3924 (1984).
Wilkins, J. A. & Lin, S. J. Cell Biol. 102, 1085–1092 (1986).
Ruhnau, K. & Wegner, A. FEBS Lett. 228, 105–108 (1988).
Westmeyer, A., Ruhnau, K., Wegner, A. & Jockusch, B. M. EMBO J. 9, 2071–2078 (1990).
Otto, J. J. Cell Motil. Cytoskel. 6, 48–55 (1986).
Luna, E. J. & Hitt, A. L. Science 258, 955–964 (1992).
Johnson, R. P. & Craig, S. W. J. biol. Chem. 269, 12611–12619 (1994).
Molony, L. & Burridge, K. J. Cell Biochem. 29, 31–36 (1985).
Coutu, M. D. & Craig, S. W. Proc. natn. Acad. Sci. U.S.A. 85, 8535–8539 (1988).
Price, G. J. et al. Biochem. J. 259, 453–461 (1989).
Menkel, A. R., Ronsiek, M., Nikolai, G. & Jockusch, B. M. Eur. J. Cell Biol. (abstr.) 60 (suppl. 37), 38 (1993).
Menkel, A. R. et al. J. Cell Biol. 126, 1231–1240 (1994).
MacLean-Fletcher, S. D. & Pollard, T. D. J. Cell Biol. 85, 414–428 (1980).
Stossel, T. P. et al. A. Rev. Cell Biol. 1, 353–402 (1985).
Gimona, M. et al. EMBO J. 7, 2329–2334 (1988).
Price, G. J. et al. Biochem. J. 245, 595–603 (1987).
Critchley, D. R. et al. Biochem. Soc. Trans. 19, 1028–1033 (1991).
Feramisco, J. R. & Burridge, K. J. biol. Chem. 255, 1194–1199 (1980).
Groesch, M. E. & Otto, J. J. Cell Motil. Cytoskel. 15, 41–50 (1990).
Smith, D. B. & Johnson, K. S. Gene 67, 31–40 (1988).
Spudich, J. & Watt, S. J. biol. Chem. 246, 4866–4871 (1971).
MacLean-Fletcher, S. & Pollard, T. D. Biochem. biophys. Res. Commun. 96, 18–27 (1980).
Laemmli, U. K. Nature 227, 680–685 (1970).
Frorath, B., Abney, C. C., Berthold, H., Scanarini, M. & Northemann, W. Biotechniques 12, 558–563 (1992).
Milam, L. M. J. molec. Biol. 134, 543–545 (1985).
Gilmore, A. P. et al. J. Cell Biol. 122, 337–347 (1993).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Johnson, R., Craig, S. F-actin binding site masked by the intramolecular association of vinculin head and tail domains. Nature 373, 261–264 (1995). https://doi.org/10.1038/373261a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/373261a0
This article is cited by
-
Vinculin-mediated axon growth requires interaction with actin but not talin in mouse neocortical neurons
Cellular and Molecular Life Sciences (2021)
-
Vinexin family (SORBS) proteins regulate mechanotransduction in mesenchymal stem cells
Scientific Reports (2018)
-
Molecular mechanism of vinculin activation and nanoscale spatial organization in focal adhesions
Nature Cell Biology (2015)
-
How far in-silico computing meets real experiments. A study on the structure and dynamics of spin labeled vinculin tail protein by molecular dynamics simulations and EPR spectroscopy
BMC Genomics (2013)
-
Dimer asymmetry defines α-catenin interactions
Nature Structural & Molecular Biology (2013)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.