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.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Scientific Reports Open Access 19 May 2022
Scientific Reports Open Access 07 March 2022
Signal Transduction and Targeted Therapy Open Access 30 August 2021
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Protein Data Bank
Cadigan, K.M. & Nusse, R. Wnt signaling: a common theme in animal development. Genes Dev. 11, 3286–3305 (1997).
Beachy, P.A., Karhadkar, S.S. & Berman, D.M. Tissue repair and stem cell renewal in carcinogenesis. Nature 432, 324–331 (2004).
Polakis, P. Wnt signaling and cancer. Genes Dev. 14, 1837–1851 (2000).
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).
Theisen, H. et al. dishevelled is required during wingless signaling to establish both cell polarity and cell identity. Development 120, 347–360 (1994).
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).
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).
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).
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).
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).
Rothbacher, U. et al. Dishevelled phosphorylation, subcellular localization and multimerization regulate its role in early embryogenesis. EMBO J. 19, 1010–1022 (2000).
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).
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).
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).
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).
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).
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).
Semenov, M.V. & Snyder, M. Human dishevelled genes constitute a DHR-containing multigene family. Genomics 42, 302–310 (1997).
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).
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).
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).
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).
Schwarz-Romond, T., Metcalfe, C. & Bienz, M. Dynamic recruitment of Axin by Dishevelled protein assemblies. J. Cell Sci. (in the press).
Capelluto, D.G. et al. The DIX domain targets dishevelled to actin stress fibres and vesicular membranes. Nature 419, 726–729 (2002).
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).
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).
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).
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).
Korinek, V. et al. Constitutive transcriptional activation by a β-catenin-Tcf complex in APC−/− colon carcinoma. Science 275, 1784–1787 (1997).
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).
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).
Serpell, L.C., Sunde, M. & Blake, C.C. The molecular basis of amyloidosis. Cell. Mol. Life Sci. 53, 871–887 (1997).
Jones, D.T. Protein secondary structure prediction based on position-specific scoring matrices. J. Mol. Biol. 292, 195–202 (1999).
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).
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).
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).
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).
Bilic, J. et al. Wnt induces LRP6 signalosomes and promotes Dishevelled-dependent LRP6 phosphorylation. Science (in the press).
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).
Sear, R. Dishevelled: a protein that functions in living cells by phase separating. Soft Matter (in the press).
Carron, C. et al. Frizzled receptor dimerization is sufficient to activate the Wnt/β-catenin pathway. J. Cell Sci. 116, 2541–2550 (2003).
Tamai, K. et al. A mechanism for Wnt coreceptor activation. Mol. Cell 13, 149–156 (2004).
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).
Kim, C.A. & Bowie, J.U. SAM domains: uniform structure, diversity of function. Trends Biochem. Sci. 28, 625–628 (2003).
Qiao, F. et al. Derepression by depolymerization; structural insights into the regulation of Yan by Mae. Cell 118, 163–173 (2004).
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).
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).
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).
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).
Philo, J.S. Improved methods for fitting sedimentation coefficient distributions derived by time-derivative techniques. Anal. Biochem. 354, 238–246 (2006).
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).
Leslie, A.G. Integration of macromolecular diffraction data. Acta Crystallogr. D Biol. Crystallogr. 55, 1696–1702 (1999).
Collaborative Computational Project, Number 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).
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).
Weeks, C.M. & Miller, R. The design and implementation of SnB v2.0. J. Appl. Cryst. 32, 120–124 (1997).
McRee, D.E. Practical Protein Crystallography (Academic Press, San Diego, 1993).
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).
Kraulis, J. MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Cryst. 24, 946–950 (1991).
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.).
The authors declare no competing financial interests.
Circular dichroism of wild-type and mutant DIX domains. (PDF 241 kb)
Additional sedimentation analysis of wild-type and mutant DIX domains. (PDF 88 kb)
The degree of Dvl2 oligomerization is not altered by Wnt signaling. (PDF 1777 kb)
The electron density peak at the molecular interface. (PDF 5687 kb)
About this article
Cite this article
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
This article is cited by
Scientific Reports (2022)
Scientific Reports (2022)
Cell Research (2021)
Nature Reviews Cancer (2021)
Signal Transduction and Targeted Therapy (2021)