Colorectal cancer results from mutations in components of the Wnt pathway that regulate β-catenin levels1. Dishevelled (Dvl or Dsh) signals downstream of Wnt receptors and stabilizes β-catenin during cell proliferation1 and embryonic axis formation2. Moreover, Dvl contributes to cytoskeletal reorganization during gastrulation3,4,5 and mitotic spindle orientation during asymmetric cell division6. Dvl belongs to a family of eukaryotic signalling proteins that contain a conserved 85-residue module of unknown structure and biological function called the DIX domain7. Here we show that the DIX domain mediates targeting to actin stress fibres and cytoplasmic vesicles in vivo. Neighbouring interaction sites for actin and phospholipid are identified between two helices by nuclear magnetic resonance spectroscopy (NMR). Mutation of the actin-binding motif abolishes the cytoskeletal localization of Dvl, but enhances Wnt/β-catenin signalling and axis induction in Xenopus. By contrast, mutation of the phospholipid interaction site disrupts vesicular association of Dvl, Dvl phosphorylation, and Wnt/β-catenin pathway activation. We propose that partitioning of Dvl into cytoskeletal and vesicular pools by the DIX domain represents a point of divergence in Wnt signalling.
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Peifer, M. & Polakis, P. Wnt signaling in oncogenesis and embryogenesis—a look outside the nucleus. Science 287, 1606–1609 (2000)
Moon, R. T. & Kimelman, D. From cortical rotation to organizer gene expression: toward a molecular explanation of axis specification in Xenopus. BioEssays 20, 536–545 (1998)
Habas, R., Kato, Y. & He, X. Wnt/Frizzled activation of Rho regulates vertebrate gastrulation and requires a novel Formin homology protein Daam1. Cell 107, 843–854 (2001)
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)
Wallingford, J. B. et al. Dishevelled controls cell polarity during Xenopus gastrulation. Nature 405, 81–85 (2000)
Gho, M. & Schweisguth, F. Frizzled signalling controls orientation of asymmetric sense organ precursor cell divisions in Drosophila. Nature 393, 178–181 (1998)
Cadigan, K. M. & Nusse, R. Wnt signaling: a common theme in animal development. Genes Dev. 11, 3286–3305 (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)
Pantaloni, D., Le Clainche, C. & Carlier, M. F. Mechanism of actin-based motility. Science 292, 1502–1506 (2001)
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)
Krylova, O., Messenger, M. J. & Salinas, P. C. Dishevelled-1 regulates microtubule stability: a new function mediated by glycogen synthase kinase-3β. J. Cell Biol. 151, 83–94 (2000)
Morfini, G., Szebenyi, G., Elluru, R., Ratner, N. & Brady, S. T. Glycogen synthase kinase 3 phosphorylates kinesin light chains and negatively regulates kinesin-based motility. EMBO J. 21, 281–293 (2002)
Adamson, P., Paterson, H. F. & Hall, A. Intracellular localization of the P21rho proteins. J. Cell Biol. 119, 617–627 (1992)
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)
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)
Sun, T. Q. et al. PAR-1 is a Dishevelled-associated kinase and a positive regulator of Wnt signalling. Nature Cell Biol. 3, 628–636 (2001)
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)
Li, L. et al. Axin and Frat1 interact with Dvl and GSK, bridging Dvl to GSK in Wnt-mediated regulation of LEF-1. EMBO J. 18, 4233–4240 (1999)
Henry, G. D. & Sykes, B. D. Methods to study membrane protein structure in solution. Methods Enzymol. 239, 515–535 (1994)
Yarmola, E. G., Edison, A. S., Lenox, R. H. & Bubb, M. R. Actin filament cross-linking by MARCKS: characterization of two actin-binding sites within the phosphorylation site domain. J. Biol. Chem. 276, 22351–22358 (2001)
Vancompernolle, K., Vandekerckhove, J., Bubb, M. R. & Korn, E. D. The interfaces of actin and Acanthamoeba actobindin. Identification of a new actin-binding motif. J. Biol. Chem. 266, 15427–15431 (1991)
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)
Rothbacher, U. et al. Dishevelled phosphorylation, subcellular localization and multimerization regulate its role in early embryogenesis. EMBO J. 19, 1010–1022 (2000)
Fagotto, F. et al. Domains of axin involved in protein-protein interactions, Wnt pathway inhibition, and intracellular localization. J. Cell Biol. 145, 741–756 (1999)
Axelrod, J. D. Unipolar membrane association of Dishevelled mediates Frizzled planar cell polarity signaling. Genes Dev. 15, 1182–1187 (2001)
Peters, J. M., McKay, R. M., McKay, J. P. & Graff, J. M. Casein kinase I transduces Wnt signals. Nature 401, 345–350 (1999)
Seeling, J. M. et al. Regulation of β-catenin signaling by the B56 subunit of protein phosphatase 2A. Science 283, 2089–2091 (1999)
Dahmen, R. P. et al. Deletions of axin1, a component of the wnt/wingless pathway, in sporadic medulloblastomas. Cancer Res. 61, 7039–7043 (2001)
Anderson, C. W., Baum, P. R. & Gesteland, R. F. Processing of adenovirus 2-induced proteins. J. Virol. 12, 241–252 (1973)
Kato, Y., Shi, Y. & He, X. Neuralization of the Xenopus embryo by inhibition of p300/CREB-binding protein function. J. Neurosci. 19, 9364–9373 (1999)
C.V.F. conceived, executed and interpreted the immunofluorescence experiments and designed and analysed the immunoprecipitation and dephosphorylation experiments. We thank X. Chen, M. L. Dell'Acqua, L. E. Heasley, R. S. Hodges, J. E. Hooper, D. N. M. Jones, A. L. Lewellyn, J. L. Maller and A. D. Sorkin for discussions and support. This work was supported by the National Institutes of Health (M.O.) and the University of Colorado Health Sciences Center's Biophysical, DNA Sequencing, and NMR Facilities. C.V.F. received support from the Howard Hughes Medical Institute as a Research Associate. R.H. is supported by a NIH training grant, and acknowledges I. Dawid for encouragement. X.H. and M.O. are Pew Scholars and X.H. is a Keck Foundation Distinguished Young Scholar.
The authors declare that they have no competing financial interests.
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Capelluto, D., Kutateladze, T., Habas, R. et al. The DIX domain targets dishevelled to actin stress fibres and vesicular membranes. Nature 419, 726–729 (2002). https://doi.org/10.1038/nature01056
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