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Wnt signalling regulates paxillin ubiquitination essential for mesodermal cell motility

Nature Cell Biology volume 9, pages 813–821 (2007)Cite this article

Abstract

Gastrulation movements are critical for establishing the three germ layers and the architecture of vertebrate embryos. During Xenopus laevis gastrulation, mesodermal tissue migrates on the blastocoel roof and elongates along the antero-posterior axis1,2. During this process, cells in the dorsal mesoderm are polarized and intercalate with each other, which is defined as convergent extension and is known to be regulated by the non-canonical Wnt pathway3,4,5. Here, we show that paxillin plays an essential role in this process. Paxillin is a focal-adhesion associated protein implicated in the regulation of actin cytoskeletal organization and cell motility6,7, but its role in Xenopus embryogenesis has not yet been clarified. We demonstrate that the Wnt pathway controls the ubiquitination and stability of paxillin, and that this regulatory mechanism is essential for convergent extension movements. We identified a RING finger protein XRNF185, which physically binds to paxillin and the proteasome. XRNF185 destabilizes paxillin at focal adhesions and promotes mesodermal cell migration during convergent extension. We propose a mechanism to regulate gastrulation movements that involves paxillin ubiquitination and stability controlled by Wnt signalling.

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Figure 1: Paxillin localizes to focal adhesion-like structures and has an essential role during Xenopus gastrulation.
Figure 2: Paxillin ubiquitination is regulated by non-canonical Wnt signalling.
Figure 3: XRNF185 regulates Paxillin stability.
Figure 4: XRNF185, 20S proteasome and paxillin were colocalized in focal adhesions.
Figure 5: Wnt pathway regulates paxillin turnover at focal adhesions through XRNF185 activity.

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References

  1. Wallingford, J. B., Fraser, S. E. & Harland, R. M. Convergent extension: the molecular control of polarized cell movement during embryonic development. Dev. Cell 2, 695–706 (2002).

    Article  CAS  Google Scholar 

  2. Leptin, M. Gastrulation movements: the logic and the nuts and bolts. Dev. Cell 8, 305–320 (2005).

    Article  CAS  Google Scholar 

  3. Heisenberg, C. P. et al. Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation. Nature 405, 76–81 (2000).

    Article  CAS  Google Scholar 

  4. Smith, J. C., Conlon, F. L., Saka, Y. & Tada, M. Xwnt11 and the regulation of gastrulation in Xenopus. Philos. Trans. R. Soc. Lond. B Biol. Sci. 355, 923–930 (2000).

    Article  CAS  Google Scholar 

  5. Wallingford, J. B. et al. Dishevelled controls cell polarity during Xenopus gastrulation. Nature 405, 81–85 (2000).

    Article  CAS  Google Scholar 

  6. Turner, C. E. Paxillin and focal adhesion signalling. Nature Cell Biol. 2, E231–E236 (2000).

    Article  CAS  Google Scholar 

  7. Carragher, N. O. & Frame, M. C. Focal adhesion and actin dynamics: a place where kinases and proteases meet to promote invasion. Trends Cell Biol. 14, 241–249 (2004).

    Article  CAS  Google Scholar 

  8. Li, M. & Sakaguchi, D. S. Inhibition of integrin-mediated adhesion and signaling disrupts retinal development. Dev. Biol 275, 202–214 (2004).

    Article  CAS  Google Scholar 

  9. Iioka, H., Ueno, N. & Kinoshita, N. Essential role of MARCKS in cortical actin dynamics during gastrulation movements. J. Cell Biol. 164, 169–174 (2004).

    Article  CAS  Google Scholar 

  10. Critchley, D. R. Cytoskeletal proteins talin and vinculin in integrin-mediated adhesion. Biochem. Soc. Trans. 32, 831–836 (2004).

    Article  CAS  Google Scholar 

  11. Ziegler, W. H., Liddington, R. C. & Critchley, D. R. The structure and regulation of vinculin. Trends Cell Biol. 16, 453–460 (2006).

    Article  CAS  Google Scholar 

  12. Didier, C. et al. RNF5, a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization. Mol. Cell Biol. 23, 5331–5345 (2003).

    Article  CAS  Google Scholar 

  13. Tada, M. & Smith, J. C. Xwnt11 is a target of Xenopus Brachyury: regulation of gastrulation movements via Dishevelled, but not through the canonical Wnt pathway. Development 127, 2227–2238 (2000).

    CAS  PubMed  Google Scholar 

  14. Gong, Y., Mo, C. & Fraser, S. E. Planar cell polarity signalling controls cell division orientation during zebrafish gastrulation. Nature 430, 689–693 (2004).

    Article  CAS  Google Scholar 

  15. Sheldahl, L. C. et al. Dishevelled activates Ca2+ flux, PKC, and CamKII in vertebrate embryos. J. Cell Biol. 161, 769–777 (2003).

    Article  CAS  Google Scholar 

  16. Rothbacher, U. et al. Dishevelled phosphorylation, subcellular localization and multimerization regulate its role in early embryogenesis. EMBO J. 19, 1010–1022 (2000).

    Article  CAS  Google Scholar 

  17. Broday, L. et al. The LIM domain protein UNC-95 is required for the assembly of muscle attachment structures and is regulated by the RING finger protein RNF-5 in C. elegans. J. Cell Biol. 165, 857–867 (2004).

    Article  CAS  Google Scholar 

  18. Natsume, T. et al. A direct nanoflow liquid chromatography-tandem mass spectrometry system for interaction proteomics. Anal. Chem. 74, 4725–4733 (2002).

    Article  CAS  Google Scholar 

  19. Tokumoto, T. & Ishikawa, K. A novel “active” form of proteasomes from Xenopus laevis ovary cytosol. Biochem. Biophys. Res. Commun. 192, 1106–1114 (1993).

    Article  CAS  Google Scholar 

  20. Verma, R., Oania, R., Graumann, J. & Deshaies, R. J. Multiubiquitin chain receptors define a layer of substrate selectivity in the ubiquitin–proteasome system. Cell 118, 99–110 (2004).

    Article  CAS  Google Scholar 

  21. Ferrell, K., Wilkinson, C. R., Dubiel, W. & Gordon, C. Regulatory subunit interactions of the 26S proteasome, a complex problem. Trends Biochem. Sci. 25, 83–88 (2000).

    Article  CAS  Google Scholar 

  22. Joazeiro, C. A. & Weissman, A. M. RING finger proteins: mediators of ubiquitin ligase activity. Cell 102, 549–552 (2000).

    Article  CAS  Google Scholar 

  23. Weissman, A. M. Themes and variations on ubiquitylation. Nature Rev. Mol. Cell Biol. 2, 169–178 (2001).

    Article  CAS  Google Scholar 

  24. Reits, E. A., Benham, A. M., Plougastel, B., Neefjes, J. & Trowsdale, J. Dynamics of proteasome distribution in living cells. EMBO J. 16, 6087–6094 (1997).

    Article  CAS  Google Scholar 

  25. Sokol, S. Y. Analysis of Dishevelled signalling pathways during Xenopus development. Curr. Biol. 6, 1456–1467 (1996).

    Article  CAS  Google Scholar 

  26. Franco, S. J. et al. Calpain-mediated proteolysis of talin regulates adhesion dynamics. Nature Cell Biol. 6, 977–983 (2004).

    Article  CAS  Google Scholar 

  27. Frank, S. R., Adelstein, M. R. & Hansen, S. H. GIT2 represses Crk- and Rac1-regulated cell spreading and Cdc42-mediated focal adhesion turnover. EMBO J. 25, 1848–1859 (2006).

    Article  CAS  Google Scholar 

  28. Wehrle-Haller, B. & Imhof, B. The inner lives of focal adhesions. Trends Cell Biol. 12, 382–389 (2002).

    Article  CAS  Google Scholar 

  29. Ulrich, F. et al. Wnt11 functions in gastrulation by controlling cell cohesion through Rab5c and E-cadherin. Dev. Cell 9, 555–564 (2005).

    Article  CAS  Google Scholar 

  30. Zohn, I. E. et al. p38 and a p38-interacting protein are critical for downregulation of E-cadherin during mouse gastrulation. Cell 125, 957–969 (2006).

    Article  CAS  Google Scholar 

  31. Kinoshita, N., Iioka, H., Miyakoshi, A. & Ueno, N. PKCδ is essential for Dishevelled function in a noncanonical Wnt pathway that regulates Xenopus convergent extension movements. Genes Dev. 17, 1663–1676 (2003).

    Article  CAS  Google Scholar 

  32. Murray, A. W. Cell cycle extracts. Methods Cell Biol. 36, 581–605 (1991).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank T. Tokumoto for the anti-proteasome antibodies. We are grateful to N. Ueno and members of his lab for advice, suggestions and help throughout the course of this work. This work was supported by a Grant-in-Aid for Scientific Research (Kakenhi) to N.K. and from the New Energy and Industrial Technology Development Organization (NEDO) to S.I. and T.N.

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Authors and Affiliations

  1. Department of Developmental Biology; National Institute for Basic Biology; Okazaki,

    Hidekazu Iioka & Noriyuki Kinoshita

  2. National Institute for Basic Biology, Aichi, 444-8585, Japan

    Hidekazu Iioka & Noriyuki Kinoshita

  3. National Institutes of Advanced Industrial Science and Technology,

    Shun-ichiro Iemura & Tohru Natsume

  4. Biological Information Research Center, Tokyo, 135-0064, Japan

    Shun-ichiro Iemura & Tohru Natsume

  5. Department of Molecular Biomechanics,

    Noriyuki Kinoshita

  6. The Graduate University for Advanced Studies, Aichi, 444-8585, Japan

    Noriyuki Kinoshita

Authors
  1. Hidekazu Iioka
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  2. Shun-ichiro Iemura
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Contributions

N.K. directed the work. H.I. and N.K. contributed to experimental planning and work. S.I. and T.N. conducted mass spectrometry analysis of XRNF185 binding proteins.

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Correspondence to Noriyuki Kinoshita.

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The authors declare no competing financial interests.

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Supplementary Figures S1, S2, S3, S4, S5, S6 and Supplementary Table (PDF 2400 kb)

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Iioka, H., Iemura, Si., Natsume, T. et al. Wnt signalling regulates paxillin ubiquitination essential for mesodermal cell motility. Nat Cell Biol 9, 813–821 (2007). https://doi.org/10.1038/ncb1607

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