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Regulation of Focal Adhesion Dynamics by Wnt5a Signaling

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Book cover Planar Cell Polarity

Part of the book series: Methods in Molecular Biology ((MIMB,volume 839))

Abstract

Wnt5a is a representative ligand that activates the β-catenin-independent pathway of Wnt signaling in mammals. This pathway might be related to planar cell polarity signaling in Drosophila. Because reliable biochemical assays to measure Wnt5a pathway activity have not yet been established, we examined whether Wnt5a signaling stimulates focal adhesion turnover in migrating cells using live immunofluorescence imaging and immunocytochemical analysis. These assays demonstrated that the Wnt5a pathway cooperates with integrin signaling to regulate cell migration and adhesion through focal adhesion dynamics.

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References

  1. Adler, P. N. (2002) Planar signaling and morphogenesis in Drosophila. Dev Cell 2, 525–35.

    Article  PubMed  CAS  Google Scholar 

  2. Axelrod, J. D. (2009) Progress and challenges in understanding planar cell polarity signaling. Semin Cell Dev Biol 20, 964–71.

    Article  PubMed  CAS  Google Scholar 

  3. Veeman, M. T., Axelrod, J. D., and Moon, R. T. (2003) A second canon. Functions and mechanisms of β-catenin-independent Wnt signaling. Dev Cell 5, 367–77.

    Article  PubMed  CAS  Google Scholar 

  4. Wu, J., and Mlodzik, M. (2009) A quest for the mechanism regulating global planar cell polarity of tissues. Trends Cell Biol 19, 295–305.

    Article  PubMed  Google Scholar 

  5. Cadigan, K. M., and Nusse, R. (1997) Wnt signaling: a common theme in animal development. Genes Dev 11, 3286–305.

    Article  PubMed  CAS  Google Scholar 

  6. Logan, C. Y., and Nusse, R. (2004) The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol 20, 781–810.

    Article  PubMed  CAS  Google Scholar 

  7. Kikuchi, A., Yamamoto, H., and Sato, A. (2009) Selective activation mechanisms of Wnt signaling pathways. Trends Cell Biol 19, 119–29.

    Article  PubMed  CAS  Google Scholar 

  8. Strutt, D. I., Weber, U., and Mlodzik, M. (1997) The role of RhoA in tissue polarity and Frizzled signalling. Nature 387, 292–5.

    Article  PubMed  CAS  Google Scholar 

  9. Winter, C. G., Wang, B., Ballew, A., et al. (2001) Drosophila Rho-associated kinase (Drok) links Frizzled-mediated planar cell polarity signaling to the actin cytoskeleton. Cell 105, 81–91.

    Article  PubMed  CAS  Google Scholar 

  10. Wharton, K. A. J. (2003) Runnin’ with the Dvl: proteins that associate with Dsh/Dvl and their significance to Wnt signal transduction. Dev Biol 253, 1–17.

    Article  PubMed  CAS  Google Scholar 

  11. Du, S. J., Purcell, S. M., Christian, J. L., McGrew, L. L., and Moon, R. T. (1995) Identification of distinct classes and functional domains of Wnts through expression of wild-type and chimeric proteins in Xenopus embryos. Mol Cell Biol 15, 2625–34.

    PubMed  CAS  Google Scholar 

  12. Moon, R. T., Campbell, R. M., Christian, J. L., McGrew, L. L., Shih, J., and Fraser, S. (1993) Xwnt-5A: a maternal Wnt that affects morphogenetic movements after overexpression in embryos of Xenopus laevis. Development 119, 97–111.

    PubMed  CAS  Google Scholar 

  13. Heisenberg, C. P., Tada, M., Rauch, G. J., et al. (2000) Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation. Nature 405, 76–81.

    Article  PubMed  CAS  Google Scholar 

  14. Rauch, G. J., Hammerschmidt, M., Blader, P., et al. (1997) Wnt5 is required for tail formation in the zebrafish embryo. Cold Spring Harb Symp Quant Biol 62, 227–34.

    PubMed  CAS  Google Scholar 

  15. Kikuchi, A., and Yamamoto, H. (2008) Tumor formation due to abnormalities in the β-catenin-independent pathway of Wnt signaling. Cancer Sci 99, 202–8.

    Article  PubMed  CAS  Google Scholar 

  16. Kurayoshi, M., Oue, N., Yamamoto, H., et al. (2006) Expression of Wnt-5a is correlated with aggressiveness of gastric cancer by stimulating cell migration and invasion. Cancer Res 66, 10439–48.

    Article  PubMed  CAS  Google Scholar 

  17. Kurayoshi, M., Yamamoto, H., Izumi, S., and Kikuchi, A. (2007) Post-translational palmitoylation and glycosylation of Wnt-5a are necessary for its signalling. Biochem J 402, 515–23.

    Article  PubMed  CAS  Google Scholar 

  18. Yamamoto, H., Kitadai, Y., Oue, N., Ohdan, H., Yasui, W., and Kikuchi, A. (2009) Laminin γ2 mediates Wnt5a-induced invasion of gastric cancer cells. Gastroenterology 137, 242–52, 52 e1–6.

    Google Scholar 

  19. Matsumoto, S., Fumoto, K., Okamoto, T., Kaibuchi, K., and Kikuchi, A. (2010) Binding of APC and dishevelled mediates Wnt5a-regulated focal adhesion dynamics in migrating cells. Embo J 29, 1192–204.

    Article  PubMed  CAS  Google Scholar 

  20. Small, J. V., and Kaverina, I. (2003) Microtubules meet substrate adhesions to arrange cell polarity. Curr Opin Cell Biol 15, 40–7.

    Article  PubMed  CAS  Google Scholar 

  21. Mitra, S. K., Hanson, D. A., and Schlaepfer, D. D. (2005) Focal adhesion kinase: in command and control of cell motility. Nat Rev Mol Cell Biol 6, 56–68.

    Article  PubMed  CAS  Google Scholar 

  22. Carragher, N. O., and Frame, M. C. (2004) Focal adhesion and actin dynamics: a place where kinases and proteases meet to promote invasion. Trends Cell Biol 14, 241–9.

    Article  PubMed  CAS  Google Scholar 

  23. Ridley, A. J., Schwartz, M. A., Burridge, K., et al. (2003) Cell migration: integrating signals from front to back. Science 302, 1704–9.

    Article  PubMed  CAS  Google Scholar 

  24. Legate, K. R., Wickstrom, S. A., and Fassler, R. (2009) Genetic and cell biological analysis of integrin outside-in signaling. Genes Dev 23, 397–418.

    Article  PubMed  CAS  Google Scholar 

  25. Wozniak, M. A., Modzelewska, K., Kwong, L., and Keely, P. J. (2004) Focal adhesion regulation of cell behavior. Biochim Biophys Acta 1692, 103–19.

    Article  PubMed  CAS  Google Scholar 

  26. Schaller, M. D., Hildebrand, J. D., Shannon, J. D., Fox, J. W., Vines, R. R., and Parsons, J. T. (1994) Autophosphorylation of the focal adhesion kinase, pp125FAK, directs SH2-dependent binding of pp60src. Mol Cell Biol 14, 1680–8.

    PubMed  CAS  Google Scholar 

  27. Bellis, S. L., Perrotta, J. A., Curtis, M. S., and Turner, C. E. (1997) Adhesion of fibroblasts to fibronectin stimulates both serine and tyrosine phosphorylation of paxillin. Biochem J 325 (Pt 2), 375–81.

    PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Nature Publishing Group and Portland Press Ltd for permitting the use of original figures. Financial support was provided by Grants-in-Aid for Scientific Research and for Scientific Research on Priority Areas from the Ministry of Education, Science, and Culture of Japan (2008, 2009, 2010), and by Research Grants from the Princess Takamatsu Cancer Research Fund (08-24005) and Takeda Science Foundation (2009).

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Authors and Affiliations

  1. Department of Molecular Biology & Biochemistry, Graduate School of Medicine, Faculty of Medicine, Osaka University, Osaka, Japan

    Shinji Matsumoto & Akira Kikuchi

Authors
  1. Shinji Matsumoto
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  2. Akira Kikuchi
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Corresponding author

Correspondence to Akira Kikuchi .

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Editors and Affiliations

  1. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Smyth Road 501, Ottawa, K1Y 8L6, Canada

    Kursad Turksen

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Matsumoto, S., Kikuchi, A. (2012). Regulation of Focal Adhesion Dynamics by Wnt5a Signaling. In: Turksen, K. (eds) Planar Cell Polarity. Methods in Molecular Biology, vol 839. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-510-7_17

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  • DOI: https://doi.org/10.1007/978-1-61779-510-7_17

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-509-1

  • Online ISBN: 978-1-61779-510-7

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