Abstract
Primary cilia and basal bodies are evolutionarily conserved organelles that mediate communication between the intracellular and extracellular environments. Here we show that bbs1, bbs4 and mkks (also known as bbs6), which encode basal body proteins, are required for convergence and extension in zebrafish and interact with wnt11 and wnt5b. Suppression of bbs1, bbs4 and mkks transcripts results in stabilization of β-catenin with concomitant upregulation of T-cell factor (TCF)-dependent transcription in both zebrafish embryos and mammalian ciliated cells, a defect phenocopied by the silencing of the axonemal kinesin subunit KIF3A but not by chemical disruption of the cytoplasmic microtubule network. These observations are attributable partly to defective degradation by the proteasome; suppression of BBS4 leads to perturbed proteasomal targeting and concomitant accumulation of cytoplasmic β-catenin. Cumulatively, our data indicate that the basal body is an important regulator of Wnt signal interpretation through selective proteolysis and suggest that defects in this system may contribute to phenotypes pathognomonic of human ciliopathies.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Logan, C.Y. & Nusse, R. The Wnt signaling pathway in development and disease. Annu. Rev. Cell Dev. Biol. 20, 781–810 (2004).
Veeman, M.T., Axelrod, J.D. & Moon, R.T. A second canon: functions and mechanisms of beta-catenin-independent Wnt signaling. Dev. Cell 5, 367–377 (2003).
Aberle, H., Bauer, A., Stappert, J., Kispert, A. & Kemler, R. beta-catenin is a target for the ubiquitin-proteasome pathway. EMBO J. 16, 3797–3804 (1997).
Yan, D. et al. Elevated expression of axin2 and hnkd mRNA provides evidence that Wnt/beta-catenin signaling is elevated in human colon tumors. Proc. Natl. Acad. Sci. USA 98, 14973–14978 (2001).
Jho, E.H. et al. Wnt/beta-catenin/Tcf signaling induces the transcription of Axin2, a negative regulator of the signaling pathway. Mol. Cell. Biol. 22, 1172–1183 (2002).
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).
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).
Keller, R. Shaping the vertebrate body plan by polarized embryonic cell movements. Science 298, 1950–1954 (2002).
Torban, E., Kor, C. & Gros, P. Van Gogh-like2 (Strabismus) and its role in planar cell polarity and convergent extension in vertebrates. Trends Genet. 20, 570–577 (2004).
Curtin, J.A. et al. Mutation of Celsr1 disrupts planar polarity of inner ear hair cells and causes severe neural tube defects in the mouse. Curr. Biol. 13, 1129–1133 (2003).
Greene, N.D.E., Gerrelli, D., Van Straaten, H.W.M. & Copp, A.J. Abnormalities of floor plate, notochord and somite differentiation in the loop-tail (Lp) mouse: a model of severe neural tube defects. Mech. Dev. 73, 59–72 (1998).
Kibar, Z. et al. Ltap, a mammalian homolog of Drosophila Strabismus/Van Gogh, is altered in the mouse neural tube mutant loop-tail. Nat. Genet. 28, 251–255 (2001).
Kibar, Z. et al. Identification of a new chemically induced allele (Lp(m1Jus)) at the loop-tail locus: Morphology, histology, and genetic mapping. Genomics 72, 331–337 (2001).
Montcouquiol, M. et al. Identification of Vangl2 and Scrb1 as planar polarity genes in mammals. Nature 423, 173–177 (2003).
Huangfu, D. et al. Hedgehog signalling in the mouse requires intraflagellar transport proteins. Nature 426, 83–87 (2003).
Corbit, K.C. et al. Vertebrate Smoothened functions at the primary cilium. Nature 437, 1018–1021 (2005).
Liu, A.M., Wang, B.L. & Niswander, L.A. Mouse intraflagellar transport proteins regulate both the activator and repressor functions of Gli transcription factors. Development 132, 3103–3111 (2005).
Ross, A.J. et al. Disruption of Bardet-Biedl syndrome ciliary proteins perturbs planar cell polarity in vertebrates. Nat. Genet. 37, 1135–1140 (2005).
Badano, J.L. et al. Dissection of epistasis in oligogenic Bardet-Biedl syndrome. Nature 439, 326–330 (2006).
Marlow, F., Gonzalez, E.M., Yin, C.Y., Rojo, C. & Solnica-Krezel, L. No tail co-operates with non-canonical Wnt signaling to regulate posterior body morphogenesis in zebrafish. Development 131, 203–216 (2004).
Heisenberg, C.P. et al. Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation. Nature 405, 76–81 (2000).
Ishitani, T. et al. The TAK1-NLK-MAPK-related pathway antagonizes signalling between β-catenin and transcription factor TCF. Nature 399, 798–802 (1999).
Weidinger, G., Thorpe, C., Wuennenberg-Stapleton, K., Ngai, J. & Moon, R. The Sp1-related transcription factors sp5 and sp5-like act downstream of Wnt/beta-catenin signaling in mesoderm and neuroectoderm patterning. Curr. Biol. 15, 489–500 (2005).
Wessely, O., Agius, E., Oelgeschlager, M., Pera, E. & Robertis, E.D. Neural induction in the absence of mesoderm: beta-catenin-dependent expression of secreted BMP antagonists at the blastula stage in Xenopus. Dev. Biol. 234, 161–173 (2001).
Nojima, H. et al. Genetic evidence for involvement of maternally derived Wnt canonical signaling in dorsal determination in zebrafish. Mech. Dev. 121, 371–386 (2004).
Rosenbaum, J.L. & Child, F.M. Flagellar regeneration in protozoan flagellates. J. Cell Biol. 34, 345–364 (1967).
Veeman, M.T., Slusarski, D.C., Kaykas, A., Louie, S.H. & Moon, R.T. Zebrafish prickle, a modulator of noncanonical Wnt/Fz signaling, regulates gastrulation movements. Curr. Biol. 13, 680–685 (2003).
Kim, J.C. et al. The Bardet-Biedl protein BBS4 targets cargo to the pericentriolar region and is required for microtubule anchoring and cell cycle progression. Nat. Genet. 36, 462–470 (2004).
Korinek, V. et al. Constitutive transcriptional activation by a β-catenin–Tcf complex in APC−/− colon carcinoma. Science 275, 1784–1787 (1997).
Marszalek, J.R., Ruiz-Lozano, P., Roberts, E., Chien, K.R. & Goldstein, L.S.B. Situs inversus and embryonic ciliary morphogenesis defects in mouse mutants lacking the KIF3A subunit of kinesin-II. Proc. Natl. Acad. Sci. USA 96, 5043–5048 (1999).
Sawa, A., Khan, A.A., Hester, L.D. & Snyder, S.H. Glyceraldehyde-3-phosphate dehydrogenase: Nuclear translocation participates in neuronal and nonneuronal cell death. Proc. Natl. Acad. Sci. USA 94, 11669–11674 (1997).
Itoh, K., Brott, B.K., Bae, G.-U., Ratcliffe, M.J. & Sokol, S.Y. Nuclear localization is required for Dishevelled function in Wnt/beta-catenin signaling. J. Biol. 4, 1 (2005).
Olson, D.J. & Papkoff, J. Regulated expression of Wnt family members during proliferation of C57mg mammary cells. Cell Growth Differ. 5, 197–206 (1994).
Shimizu, H. et al. Transformation by Wnt family proteins correlates with regulation of beta-catenin. Cell Growth Differ. 8, 1349–1358 (1997).
Slusarski, D.C., Corces, V. & Moon, R. Interaction of Wnt and a Frizzled homologue triggers G-protein-linked phosphatidylinositol signalling. Nature 390, 410–413 (1997).
Chiang, A.P. et al. Homozygosity mapping with SNP arrays identifies TRIM32, an E3 ubiquitin ligase, as a Bardet-Biedl syndrome gene (BBS11). Proc. Natl. Acad. Sci. USA 103, 6287–6292 (2006).
Wigley, W.C. et al. Dynamic association of proteasomal machinery with the centrosome. J. Cell Biol. 145, 481–490 (1999).
DeMartino, G.N. Purification of PA700, the 19S regulatory subunit of the 26S proteasome. Methods Enzymol. 398, 295–306 (2005).
Deveraux, Q., Ustrell, V., Pickart, C. & Rechsteiner, M. A 26 S protease subunit that binds ubiquitin conjugates. J. Biol. Chem. 269, 7059–7061 (1994).
Deveraux, Q., Jensen, C. & Rechsteiner, M. Molecular cloning and expression of a 26S proteasome subunit enriched in dileucine repeats. J. Biol. Chem. 270, 23726–23729 (1995).
Gherman, A., Davis, E. & Katsanis, N. The ciliary proteome database: an integrated community resource for the genetic and functional dissection of cilia. Nat. Genet. 38, 961–962 (2006).
Nusse, R. Wnt signaling in disease and in development. Cell Res. 15, 28–32 (2005).
Ishitani, T. et al. The TAK1-NLK mitogen-activated protein kinase cascade functions in the Wnt-5a/Ca2+ pathway to antagonize Wnt/beta-catenin signaling. Mol. Cell. Biol. 23, 131–139 (2003).
Liang, H. et al. Wnt5a inhibits B cell proliferation and functions as a tumor suppressor in hematopoietic tissue. Cancer Cell 4, 349–360 (2003).
Mikels, A.J. & Nusse, R. Purified Wnt5a protein activates or inhibits beta-catenin–TCF signaling depending on receptor context. PLoS Biol. 4, e115 (2006).
Topol, L. et al. Wnt-5a inhibits the canonical Wnt pathway by promoting GSK-3-independent beta-catenin degradation. J. Cell Biol. 162, 899–908 (2003).
Simons, M. et al. Inversin, the gene product mutated in nephronophthisis type II, functions as a molecular switch between Wnt signalling pathways. Nat. Genet. 37, 537–543 (2005).
Davis, E.E., Brueckner, M. & Katsanis, N. The emerging complexity of the vertebrate cilium: new functional roles for an ancient organelle. Dev. Cell 11, 9–19 (2006).
Thisse, C., Thisse, B., Schilling, T.F. & Postlethwait, J.H. Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos. Development 119, 1203–1215 (1993).
Willert, K. et al. Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature 423, 448–452 (2003).
Acknowledgements
We apologize to our colleagues whose work we were unable to cite because of a strict 50-reference limit. We thank J. Nathans, J. Axelrod, L. Menezes, G. Germino and E. Davis for their critical evaluation of this manuscript, and A. Gherman for the quantification of the western blots. We also thank J. Nathans for the gift of the 293T luciferase reporter cell line, J. Kitajewski for the mouse Wnt cDNAs, G. Walz for the gift of the Myr/Pal Dvl construct, B. Yoder for the gift of anti-polaris/IFT88 antibody and S. Leach for the validated β-catenin antibody. This work was supported by grants from the German Academic Exchange Service (J.G.), the Polycystic Kidney Disease Foundation (J.B.), the National Institute of Child Health and Development (N.K.), the National Institute of Diabetes, Digestive and Kidney disorders (N.K.), the National Institute for Arthritis and Musculoskeletal disorders (S.F.) and the Medical Research Council (P.L.B.). P.L.B. is a Senior Wellcome Trust Fellow. P.A.B. is an Investigator of the Howard Hughes Medical Institute.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–6 (PDF 826 kb)
Rights and permissions
About this article
Cite this article
Gerdes, J., Liu, Y., Zaghloul, N. et al. Disruption of the basal body compromises proteasomal function and perturbs intracellular Wnt response. Nat Genet 39, 1350–1360 (2007). https://doi.org/10.1038/ng.2007.12
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ng.2007.12
This article is cited by
-
The implication of ciliary signaling pathways for epithelial–mesenchymal transition
Molecular and Cellular Biochemistry (2023)
-
Aggresome assembly at the centrosome is driven by CP110–CEP97–CEP290 and centriolar satellites
Nature Cell Biology (2022)
-
Bardet-Biedl syndrome proteins modulate the release of bioactive extracellular vesicles
Nature Communications (2021)
-
PCM1 is necessary for focal ciliary integrity and is a candidate for severe schizophrenia
Nature Communications (2020)
-
KIF3A regulates the Wnt/β-catenin pathway via transporting β-catenin during spermatogenesis in Eriocheir sinensis
Cell and Tissue Research (2020)