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Subcellular spatial regulation of canonical Wnt signalling at the primary cilium

Nature Cell Biology volume 13, pages 700–707 (2011)Cite this article

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Abstract

Mechanisms of signal transduction regulation remain a fundamental question in a variety of biological processes and diseases. Previous evidence indicates that the primary cilium can act as a signalling hub1, but its exact role in many of the described pathways has remained elusive. Here, we investigate the mechanism of cilia-mediated regulation of the canonical Wnt pathway. We found that primary cilia dampen canonical Wnt signalling through a spatial mechanism involving compartmentalization of signalling components. The cilium, through regulated intraflagellar transport, diverts Jouberin (Jbn), a ciliopathy protein and context-specific Wnt pathway regulator2, away from the nucleus and limits β-catenin nuclear entry. This repressive regulation does not silence the pathway, but instead maintains a discrete range of Wnt responsiveness; cells without cilia have potentiated Wnt responses, whereas cells with multiple cilia have inhibited responses. Furthermore, we show that this regulation occurs during embryonic development and is disrupted in cancer cell proliferation. Together these data explain a spatial mechanism of Wnt signalling regulation that may provide insight into ciliary regulation of other signalling pathways.

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Figure 1: The primary cilium dampens Wnt activity by regulating β-catenin.
Figure 2: The primary cilium inhibits Jbn-mediated Wnt pathway regulation.
Figure 3: The primary cilium sequesters Jbn and β-catenin away from the nucleus.
Figure 4: Wnt is regulated by the primary cilium during embryonic development.
Figure 5: Regulation of cancer cell proliferation by cilia-localized Jbn.

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Acknowledgements

We are grateful to members of the Gleeson laboratory for technical expertise and feedback and the UCSD Neuroscience Microscopy Core. We would like to thank C. Kintner for helpful feedback on the manuscript. We also thank K. Willert (School of Medicine, University of California, San Diego) for reagents and technical expertise. We are grateful to S. Piccolo (Departments of Histology, Microbiology and Medical Biotechnologies, University of Padua, Italy) for the BATgal mice and to A. S. Peterson (Ernest Gallo Clinic and Research Center, Emeryville, California) for the Dnchc2 mutant mice and MEFs as well as L. S. B. Goldstein (School of Medicine, University of California, San Diego) for Kif3a mutant mice. We also thank M. G. Rosenfeld (School of Medicine, University of California, San Diego) for the β-catΔN construct and R. T. Moon (Department of Pharmacology, University of Washington) for the Super Topflash construct, as well as T. Caspary (Department of Human Genetics, Emory University School of Medicine) for the Arl13b antibody, C. Janke (Curie Institut, Paris, France) for GT335 antibody and R. B. Vallee (Department of Pathology and Cell Biology, Columbia University) for Dnchc1 and Dnchc2 antibodies. We also thank P. Mellon (School of Medicine, University of California, San Diego) for the β-galactosidase expression construct for luciferase assays. M.A.L. received support from the Bear Necessities Pediatric Cancer Foundation. This work was supported by the US National Institutes of Health, and the Burroughs Wellcome Fund in Translational Research (J.G.G.). J.G.G. is an investigator with Howard Hughes Medical Institute.

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  1. Madeline A. Lancaster

    Present address: Present address: IMBA—Institute of Molecular Biotechnology of the Austrian Academy of Science, Vienna 1030, Austria,

Authors and Affiliations

  1. Department of Neurosciences, Laboratory for Neurogenetics, Howard Hughes Medical Institute, Biomedical Sciences Program, LBR 3A16, 9500 Gilman Drive, University of California, San Diego, La Jolla, California 92093-0691, USA

    Madeline A. Lancaster, Jana Schroth & Joseph G. Gleeson

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J.G.G. and M.A.L. conceived and designed the experimental approach, interpreted data and wrote the manuscript. M.A.L. and J.S. carried out experiments. J.G.G. directed and supervised the project.

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Correspondence to Joseph G. Gleeson.

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Lancaster, M., Schroth, J. & Gleeson, J. Subcellular spatial regulation of canonical Wnt signalling at the primary cilium. Nat Cell Biol 13, 700–707 (2011). https://doi.org/10.1038/ncb2259

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