Skip to main content
  • Short communication
  • Published:

Zyxin-VASP interactions alter actin regulatory activity in zyxin-VASP complexes

Abstract

Cell-cell and cell-substrate adhesions are sites of dramatic actin rearrangements and where actin-membrane connections are tightly regulated. Zyxin-VASP complexes localize to sites of cell-cell and cell-substrate adhesion and function to regulate actin dynamics and actin-membrane connections at these sites. To accomplish these functions, zyxin recruits VASP to cellular sites via proline-rich binding sites near zyxin’s amino terminus. While the prevailing thought has been that zyxin simply acts as a scaffold protein for VASP binding, the identification of a LIM domain-VASP interaction could complicate this view. Here we assess how zyxin-VASP binding through both the proline rich motifs and the LIM domains alters specific VASP functions. We find that neither individual interaction alters VASP’s actin regulatory activities. In contrast, however, we find that full-length zyxin dramatically reduces VASPmediated actin bundling and actin assembly. Taken together, these results suggest a model where zyxin-VASP complexes occur in complex organizations with suppressed actin regulatory activity.

Abbreviations

VASP:

vasodilator stimulated phosphoprotein

References

  1. Critchley, D.R. Focal adhesions — the cytoskeletal connection. Curr. Opin. Cell Biol. 12 (2000) 133–139.

    Article  PubMed  CAS  Google Scholar 

  2. Wiesner, S., Legate, K.R. and Fassler, R. Integrin-actin interactions. Cell Mol. Life Sci. 62 (2005) 1081–1099.

    Article  PubMed  CAS  Google Scholar 

  3. Adams, C.L., Nelson, W.J. and Smith, S.J. Quantitative analysis of cadherincateninactin reorganization during development of cell-cell adhesion. J. Cell Biol. 135 (1996) 1899–1911.

    Article  PubMed  CAS  Google Scholar 

  4. Hansen, M.D. and Beckerle, M.C. Opposing roles of zyxin/LPP ACTA repeats and the LIM domain region in cell-cell adhesion. J. Biol. Chem. 281 (2006) 16178–16188.

    Article  PubMed  CAS  Google Scholar 

  5. Vasioukhin, V., Bauer, C., Yin, M. and Fuchs, E. Directed actin polymerization is the driving force for epithelial cell-cell adhesion. Cell 100 (2000) 209–219.

    Article  PubMed  CAS  Google Scholar 

  6. Crawford, A.W. and Beckerle, M.C. Purification and characterization of zyxin, an 82,000-dalton component of adherens junctions. J. Biol. Chem. 266 (1991) 5847–5853.

    PubMed  CAS  Google Scholar 

  7. Reinhard, M., Rudiger, M., Jockusch, B.M. and Walter, U. VASP interaction with vinculin: a recurring theme of interactions with proline-rich motifs. FEBS Lett. 399 (1996) 103–107.

    Article  PubMed  CAS  Google Scholar 

  8. Drees, B., Friederich, E., Fradelizi, J., Louvard, D., Beckerle, M.C. and Golsteyn, R.M. Characterization of the interaction between zyxin and members of the Ena/vasodilator-stimulated phosphoprotein family of proteins. J. Biol. Chem. 275 (2000) 22503–22511.

    Article  PubMed  CAS  Google Scholar 

  9. Niebuhr, K., Ebel, F., Frank, R., Reinhard, M., Domann, E., Carl, U.D., Walter, U., Gertler, F.B., Wehland, J. and Chakraborty, T. A novel prolinerich motif present in ActA of Listeria monocytogenes and cytoskeletal proteins is the ligand for the EVH1 domain, a protein module present in the Ena/VASP family. EMBO J. 16 (1997) 5433–5444.

    Article  PubMed  CAS  Google Scholar 

  10. Moody, J.D., Grange, J., Ascione, M.P., Boothe, D., Bushnell, E. and Hansen, M.D. A zyxin head-tail interaction regulates zyxin-VASP complex formation. Biochem. Biophys. Res Commun. 378 (2009) 625–628.

    Article  PubMed  CAS  Google Scholar 

  11. Hoffman, L.M., Jensen, C.C., Kloeker, S., Wang, C.L., Yoshigi, M. and Beckerle, M.C. Genetic ablation of zyxin causes Mena/VASP mislocalization, increased motility, and deficits in actin remodeling. J. Cell Biol. 172 (2006) 771–782.

    Article  PubMed  CAS  Google Scholar 

  12. Drees, B.E., Andrews, K.M. and Beckerle, M.C. Molecular dissection of zyxin function reveals its involvement in cell motility. J. Cell Biol. 147 (1999) 1549–1560.

    Article  PubMed  CAS  Google Scholar 

  13. Yoshigi, M., Hoffman, L.M., Jensen, C.C., Yost, H.J. and Beckerle, M.C. Mechanical force mobilizes zyxin from focal adhesions to actin filaments and regulates cytoskeletal reinforcement. J. Cell Biol. 171 (2005) 209–215.

    Article  PubMed  CAS  Google Scholar 

  14. Fradelizi, J., Noireaux, V., Plastino, J., Menichi, B., Louvard, D., Sykes, C., Golsteyn, R.M. and Friederich, E. ActA and human zyxin harbour Arp2/3-independent actin-polymerization activity. Nat. Cell Biol. 3 (2001) 699–707.

    Article  PubMed  CAS  Google Scholar 

  15. Sperry, R.B., Bishop, N.H., Bramwell, J.J., Brodeur, M.N., Carter, M.J., Fowler, B.T., Lewis, Z.B. Maxfield, S.D., Staley, D.M., Vellinga, R.M. and Hansen, M.D. Zyxin controls migration in epithelial-mesenchymal transition by mediating actin-membrane linkages at cell-cell junctions. J. Cell Physiol. 222 (2010) 612–624.

    PubMed  CAS  Google Scholar 

  16. Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72 (1976) 248–254.

    Article  PubMed  CAS  Google Scholar 

  17. Mullins, R.D. and Machesky, L.M. Actin assembly mediated by Arp2/3 complex and WASP family proteins. Methods Enzymol. 325 (2000) 214–237.

    Article  PubMed  CAS  Google Scholar 

  18. Bachmann, C., Fischer, L., Walter, U. and Reinhard, M. The EVH2 domain of the vasodilator-stimulated phosphoprotein mediates tetramerization, F-actin binding, and actin bundle formation. J. Biol. Chem. 274 (1999) 23549–23557.

    Article  PubMed  CAS  Google Scholar 

  19. Barzik, M., Kotova, T.I., Higgs, H.N., Hazelwood, L., Hanein, D., Gertler, F.B. and Schafer, D.A. Ena/VASP proteins enhance actin polymerization in the presence of barbed end capping proteins. J. Biol. Chem. 280 (2005) 28653–28662.

    Article  PubMed  CAS  Google Scholar 

  20. Bear, J.E., Svitkina, T.M., Krause, M., Schafer, D.A., Loureiro, J.J., Strasser, G.A., Maly, I.V., Chaga, O.Y., Cooper, J.A., Borisy, G.G. and Gertler, F.B. Antagonism between Ena/VASP proteins and actin filament capping regulates fibroblast motility. Cell 109 (2002) 509–521.

    Article  PubMed  CAS  Google Scholar 

  21. Huttelmaier, S., Harbeck, B., Steffens, O., Messerschmidt, T., Illenberger, S. and Jockusch, B.M. Characterization of the actin binding properties of the vasodilator-stimulated phosphoprotein VASP. FEBS Lett. 451 (1999) 68–74.

    Article  PubMed  CAS  Google Scholar 

  22. Chan, C.B., Liu, X. Tang, X., Fu, H. and Ye, K. Akt phosphorylation of zyxin mediates its interaction with acinus-S and prevents acinus-triggered chromatin condensation. Cell Death Differ. 14 (2007) 1688–1699.

    Article  PubMed  CAS  Google Scholar 

  23. Call, G.S., Chung, J.Y., Davis, J.A., Price, B.D., Primavera, T.S., Thomson, N.C., Wagner, M.V. and Hansen, M.D. Zyxin phosphorylation at serine 142 modulates the zyxin head-tail interaction to alter cell-cell adhesion. Biochem. Biophys. Res. Commun. 404 (2011) 780–784.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Marc D. H. Hansen.

Additional information

These authors contributed equally to this study

Rights and permissions

Reprints and permissions

About this article

Cite this article

Grange, J., Moody, J.D., Ascione, M.P.A. et al. Zyxin-VASP interactions alter actin regulatory activity in zyxin-VASP complexes. Cell Mol Biol Lett 18, 1–10 (2013). https://doi.org/10.2478/s11658-012-0035-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-012-0035-2

Key words