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The DIX domain of Dishevelled confers Wnt signaling by dynamic polymerization

Abstract

The Wnt signaling pathway controls numerous cell fates in animal development and is also a major cancer pathway. Dishevelled (Dvl) transduces the Wnt signal by interacting with the cytoplasmic Axin complex. Dvl and Axin each contain a DIX domain whose molecular properties and structure are unknown. Here, we demonstrate that the DIX domain of Dvl2 mediates dynamic polymerization, which is essential for the signaling activity of Dvl2. The purified domain polymerizes gradually, reversibly and in a concentration dependent manner, ultimately forming fibrils. The Axin DIX domain has a novel structural fold largely composed of β-strands that engage in head-to-tail self-interaction to form filaments in the crystal. The DIX domain thus seems to mediate the formation of a dynamic interaction platform with a high local concentration of binding sites for transient Wnt signaling partners; this represents a previously uncharacterized mechanistic principle, signaling by reversible polymerization.

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Figure 1: Signaling-defective DIX domain mutants of Dvl2.
Figure 2: Self-interaction of the DIX domain.
Figure 3: Dimerization of Dvl2 is insufficient for signaling.
Figure 4: Reversible polymerization by purified DIX domain.
Figure 5: Structure of the Axin DIX domain.
Figure 6: The helical filament formed by the Axin DIX domain in the crystal.

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References

  1. Cadigan, K.M. & Nusse, R. Wnt signaling: a common theme in animal development. Genes Dev. 11, 3286–3305 (1997).

    Article  CAS  Google Scholar 

  2. Beachy, P.A., Karhadkar, S.S. & Berman, D.M. Tissue repair and stem cell renewal in carcinogenesis. Nature 432, 324–331 (2004).

    Article  CAS  Google Scholar 

  3. Polakis, P. Wnt signaling and cancer. Genes Dev. 14, 1837–1851 (2000).

    CAS  PubMed  Google Scholar 

  4. Klingensmith, J., Nusse, R. & Perrimon, N. The Drosophila segment polarity gene dishevelled encodes a novel protein required for response to the wingless signal. Genes Dev. 8, 118–130 (1994).

    Article  CAS  Google Scholar 

  5. Theisen, H. et al. dishevelled is required during wingless signaling to establish both cell polarity and cell identity. Development 120, 347–360 (1994).

    CAS  PubMed  Google Scholar 

  6. Yanagawa, S., van Leeuwen, F., Wodarz, A., Klingensmith, J. & Nusse, R. The dishevelled protein is modified by wingless signaling in Drosophila. Genes Dev. 9, 1087–1097 (1995).

    Article  CAS  Google Scholar 

  7. Yang-Snyder, J., Miller, J.R., Brown, J.D., Lai, C.J. & Moon, R.T. A frizzled homolog functions in a vertebrate Wnt signaling pathway. Curr. Biol. 6, 1302–1306 (1996).

    Article  CAS  Google Scholar 

  8. Axelrod, J.D., Miller, J.R., Shulman, J.M., Moon, R.T. & Perrimon, N. Differential recruitment of Dishevelled provides signaling specificity in the planar cell polarity and Wingless signaling pathways. Genes Dev. 12, 2610–2622 (1998).

    Article  CAS  Google Scholar 

  9. Miller, J.R. et al. Establishment of the dorsal-ventral axis in Xenopus embryos coincides with the dorsal enrichment of dishevelled that is dependent on cortical rotation. J. Cell Biol. 146, 427–437 (1999).

    Article  CAS  Google Scholar 

  10. Umbhauer, M. et al. The C-terminal cytoplasmic Lys-thr-X-X-X-Trp motif in frizzled receptors mediates Wnt/β-catenin signalling. EMBO J. 19, 4944–4954 (2000).

    Article  CAS  Google Scholar 

  11. 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 

  12. Cong, F., Schweizer, L. & Varmus, H. Wnt signals across the plasma membrane to activate the β-catenin pathway by forming oligomers containing its receptors, Frizzled and LRP. Development 131, 5103–5115 (2004).

    Article  CAS  Google Scholar 

  13. Wong, H.C. et al. Direct binding of the PDZ domain of Dishevelled to a conserved internal sequence in the C-terminal region of Frizzled. Mol. Cell 12, 1251–1260 (2003).

    Article  CAS  Google Scholar 

  14. Kishida, S. et al. DIX domains of dvl and axin are necessary for protein interactions and their ability to regulate β-catenin stability. Mol. Cell. Biol. 19, 4414–4422 (1999).

    Article  CAS  Google Scholar 

  15. Smalley, M.J. et al. Interaction of axin and dvl-2 proteins regulates dvl-2-stimulated TCF-dependent transcription. EMBO J. 18, 2823–2835 (1999).

    Article  CAS  Google Scholar 

  16. Penton, A., Wodarz, A. & Nusse, R. A mutational analysis of dishevelled in Drosophila defines novel domains in the dishevelled protein as well as novel suppressing alleles of axin. Genetics 161, 747–762 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Cliffe, A., Hamada, F. & Bienz, M. A role of Dishevelled in relocating Axin to the plasma membrane during Wingless signaling. Curr. Biol. 13, 960–966 (2003).

    Article  CAS  Google Scholar 

  18. Semenov, M.V. & Snyder, M. Human dishevelled genes constitute a DHR-containing multigene family. Genomics 42, 302–310 (1997).

    Article  CAS  Google Scholar 

  19. Torres, M.A. & Nelson, W.J. Colocalization and redistribution of dishevelled and actin during Wnt-induced mesenchymal morphogenesis. J. Cell Biol. 149, 1433–1442 (2000).

    Article  CAS  Google Scholar 

  20. Hawkins, N.C., Ellis, G.C., Bowerman, B. & Garriga, G. MOM-5 Frizzled regulates the distribution of DSH-2 to control C. elegans asymmetric neuroblast divisions. Dev. Biol. 284, 246–259 (2005).

    Article  CAS  Google Scholar 

  21. Chang, W., Lloyd, C.E. & Zarkower, D. DSH-2 regulates asymmetric cell division in the early C. elegans somatic gonad. Mech. Dev. 122, 781–789 (2005).

    Article  CAS  Google Scholar 

  22. Itoh, K., Brott, B.K., Bae, G.U., Ratcliffe, M.J. & Sokol, S.Y. Nuclear localization is required for Dishevelled function in Wnt/β-catenin signaling. J. Biol. 4, 3 (2005).

    Article  Google Scholar 

  23. Schwarz-Romond, T., Metcalfe, C. & Bienz, M. Dynamic recruitment of Axin by Dishevelled protein assemblies. J. Cell Sci. (in the press).

  24. Capelluto, D.G. et al. The DIX domain targets dishevelled to actin stress fibres and vesicular membranes. Nature 419, 726–729 (2002).

    Article  CAS  Google Scholar 

  25. Schwarz-Romond, T., Merrifield, C., Nichols, B.J. & Bienz, M. The Wnt signalling effector Dishevelled forms dynamic protein assemblies rather than stable associations with cytoplasmic vesicles. J. Cell Sci. 118, 5269–5277 (2005).

    Article  CAS  Google Scholar 

  26. Smalley, M.J. et al. Dishevelled (Dvl-2) activates canonical Wnt signalling in the absence of cytoplasmic puncta. J. Cell Sci. 118, 5279–5289 (2005).

    Article  CAS  Google Scholar 

  27. Park, T.J., Gray, R.S., Sato, A., Habas, R. & Wallingford, J.B. Subcellular localization and signaling properties of dishevelled in developing vertebrate embryos. Curr. Biol. 15, 1039–1044 (2005).

    Article  CAS  Google Scholar 

  28. Boutros, M., Paricio, N., Strutt, D.I. & Mlodzik, M. Dishevelled activates JNK and discriminates between JNK pathways in planar polarity and wingless signaling. Cell 94, 109–118 (1998).

    Article  CAS  Google Scholar 

  29. Korinek, V. et al. Constitutive transcriptional activation by a β-catenin-Tcf complex in APC−/− colon carcinoma. Science 275, 1784–1787 (1997).

    Article  CAS  Google Scholar 

  30. Rodrigues, G.A. & Park, M. Dimerization mediated through a leucine zipper activates the oncogenic potential of the met receptor tyrosine kinase. Mol. Cell. Biol. 13, 6711–6722 (1993).

    Article  CAS  Google Scholar 

  31. Balguerie, A. et al. The sequences appended to the amyloid core region of the HET-s prion protein determine higher-order aggregate organization in vivo. J. Cell Sci. 117, 2599–2610 (2004).

    Article  CAS  Google Scholar 

  32. Serpell, L.C., Sunde, M. & Blake, C.C. The molecular basis of amyloidosis. Cell. Mol. Life Sci. 53, 871–887 (1997).

    Article  CAS  Google Scholar 

  33. Jones, D.T. Protein secondary structure prediction based on position-specific scoring matrices. J. Mol. Biol. 292, 195–202 (1999).

    Article  CAS  Google Scholar 

  34. Sakanaka, C. & Williams, L.T. Functional domains of axin. Importance of the C terminus as an oligomerization domain. J. Biol. Chem. 274, 14090–14093 (1999).

    Article  CAS  Google Scholar 

  35. Hsu, W., Zeng, L. & Costantini, F. Identification of a domain of Axin that binds to the serine/threonine protein phosphatase 2A and a self-binding domain. J. Biol. Chem. 274, 3439–3445 (1999).

    Article  CAS  Google Scholar 

  36. Luo, W. et al. Axin contains three separable domains that confer intramolecular, homodimeric, and heterodimeric interactions involved in distinct functions. J. Biol. Chem. 280, 5054–5060 (2004).

    Article  Google Scholar 

  37. Moscat, J., Diaz-Meco, M.T., Albert, A. & Campuzano, S. Cell signaling and function organized by PB1 domain interactions. Mol. Cell 23, 631–640 (2006).

    Article  CAS  Google Scholar 

  38. Bilic, J. et al. Wnt induces LRP6 signalosomes and promotes Dishevelled-dependent LRP6 phosphorylation. Science (in the press).

  39. Lee, E., Salic, A., Kruger, R., Heinrich, R. & Kirschner, M.W. The roles of APC and Axin derived from experimental and theoretical analysis of the Wnt pathway. PLoS Biol. 1, E10 (2003).

    Article  Google Scholar 

  40. Sear, R. Dishevelled: a protein that functions in living cells by phase separating. Soft Matter (in the press).

  41. Carron, C. et al. Frizzled receptor dimerization is sufficient to activate the Wnt/β-catenin pathway. J. Cell Sci. 116, 2541–2550 (2003).

    Article  CAS  Google Scholar 

  42. Tamai, K. et al. A mechanism for Wnt coreceptor activation. Mol. Cell 13, 149–156 (2004).

    Article  CAS  Google Scholar 

  43. Stefani, M. Protein misfolding and aggregation: new examples in medicine and biology of the dark side of the protein world. Biochim. Biophys. Acta 1739, 5–25 (2004).

    Article  CAS  Google Scholar 

  44. Kim, C.A. & Bowie, J.U. SAM domains: uniform structure, diversity of function. Trends Biochem. Sci. 28, 625–628 (2003).

    Article  CAS  Google Scholar 

  45. Qiao, F. et al. Derepression by depolymerization; structural insights into the regulation of Yan by Mae. Cell 118, 163–173 (2004).

    Article  CAS  Google Scholar 

  46. Bhattacharjya, S., Xu, P., Chakrapani, M., Johnston, L. & Ni, F. Polymerization of the SAM domain of MAPKKK Ste11 from the budding yeast: implications for efficient signaling through the MAPK cascades. Protein Sci. 14, 828–835 (2005).

    Article  CAS  Google Scholar 

  47. Schwarz-Romond, T. et al. The ankyrin repeat protein Diversin recruits Casein kinase Iε to the β-catenin degradation complex and acts in both canonical Wnt and Wnt/JNK signaling. Genes Dev. 16, 2073–2084 (2002).

    Article  CAS  Google Scholar 

  48. Leonard, T.A., Butler, P.J. & Löwe, J. Bacterial chromosome segregation: structure and DNA binding of the Soj dimer - a conserved biological switch. EMBO J. 24, 270–282 (2005).

    Article  CAS  Google Scholar 

  49. Cabezon, E., Butler, P.J., Runswick, M.J., Carbajo, R.J. & Walker, J.E. Homologous and heterologous inhibitory effects of ATPase inhibitor proteins on F-ATPases. J. Biol. Chem. 277, 41334–41341 (2002).

    Article  CAS  Google Scholar 

  50. Philo, J.S. Improved methods for fitting sedimentation coefficient distributions derived by time-derivative techniques. Anal. Biochem. 354, 238–246 (2006).

    Article  CAS  Google Scholar 

  51. Serpell, L.C., Berriman, J., Jakes, R., Goedert, M. & Crowther, R.A. Fiber diffraction of synthetic α-synuclein filaments shows amyloid-like cross-β conformation. Proc. Natl. Acad. Sci. USA 97, 4897–4902 (2000).

    Article  CAS  Google Scholar 

  52. Leslie, A.G. Integration of macromolecular diffraction data. Acta Crystallogr. D Biol. Crystallogr. 55, 1696–1702 (1999).

    Article  CAS  Google Scholar 

  53. Collaborative Computational Project, Number 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).

  54. de la Fortelle, E. & Bricogne, G. Maximum-likelihood heavy-atom parameter refinement for the multiple isomorphous replacement and multiwavelength anomalous diffraction methods. Methods Enzymol. 276, 472–494 (1997).

    Article  CAS  Google Scholar 

  55. Weeks, C.M. & Miller, R. The design and implementation of SnB v2.0. J. Appl. Cryst. 32, 120–124 (1997).

    Article  Google Scholar 

  56. McRee, D.E. Practical Protein Crystallography (Academic Press, San Diego, 1993).

    Google Scholar 

  57. Brunger, A.T. et al. Crystallography & NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr. D Biol. Crystallogr. 54, 905–921 (1998).

    Article  CAS  Google Scholar 

  58. Kraulis, J. MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Cryst. 24, 946–950 (1991).

    Article  Google Scholar 

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Acknowledgements

We thank P. Evans, T. Crowther and S. Li for technical help and advice, M. Semenov (Harvard Medical School) for Dvl2 antiserum, C. Niehrs for discussion and communication of unpublished results, and H. Pelham, M. Goedert, B. Nichols, J. Murphy, R. Sear, Y. Tomimoto, H. Yamamoto, H. Komori, Y. Shomura, H. Axelrod and Y. Shiro for helpful discussions. This work was supported by a European Molecular Biology Organization long-term fellowship and a Marie-Curie postdoctoral fellowship (T.S.-R.); a Erwin-Schrödinger postdoctoral fellowship (M.F.); Grants-in-Aid for Young Scientists (B) (no. 17770093) from the Japan Society for Promotion of Science and the Ministry of Education, Culture, Sports, Science and Technology, Japan (N.S.); and the 21st COE Programs, the National Project on Protein Structural and Functional Analyses (Japan) and The Japanese Aerospace Exploration Agency Project (Y.H.).

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Authors and Affiliations

Authors

Contributions

T.S.-R. conducted most of the in vivo experiments and in vitro binding studies; M.F. purified the Dvl2 DIX domain and completed the in vivo and in vitro studies; N.S. carried out the purification, crystallization and structural analysis of the Axin DIX domain; P.J.G.B. conducted the ultracentrifugation; A.K. developed the expression system for the Axin DIX domain; Y.H. directed the structural analysis of the Axin DIX domain; M.B. directed the study of Dvl2, helped with the electron microscopy and drafted the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yoshiki Higuchi or Mariann Bienz.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Circular dichroism of wild-type and mutant DIX domains. (PDF 241 kb)

Supplementary Fig. 2

Additional sedimentation analysis of wild-type and mutant DIX domains. (PDF 88 kb)

Supplementary Fig. 3

The degree of Dvl2 oligomerization is not altered by Wnt signaling. (PDF 1777 kb)

Supplementary Fig. 4

The electron density peak at the molecular interface. (PDF 5687 kb)

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Schwarz-Romond, T., Fiedler, M., Shibata, N. et al. The DIX domain of Dishevelled confers Wnt signaling by dynamic polymerization. Nat Struct Mol Biol 14, 484–492 (2007). https://doi.org/10.1038/nsmb1247

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