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Casein kinase 1 γ couples Wnt receptor activation to cytoplasmic signal transduction

Nature volume 438, pages 867–872 (2005)Cite this article

Abstract

Signalling by Wnt proteins (Wingless in Drosophila) has diverse roles during embryonic development and in adults, and is implicated in human diseases, including cancer1,2. LDL-receptor-related proteins 5 and 6 (LRP5 and LRP6; Arrow in Drosophila) are key receptors required for transmission of Wnt/β-catenin signalling in metazoa3. Although the role of these receptors in Wnt signalling is well established, their coupling with the cytoplasmic signalling apparatus remains poorly defined. Using a protein modification screen for regulators of LRP6, we describe the identification of Xenopus Casein kinase 1 γ (CK1γ), a membrane-bound member of the CK1 family. Gain-of-function and loss-of-function experiments show that CK1γ is both necessary and sufficient to transduce LRP6 signalling in vertebrates and Drosophila cells. In Xenopus embryos, CK1γ is required during anterio-posterior patterning to promote posteriorizing Wnt/β-catenin signalling. CK1γ is associated with LRP6, which has multiple, modular CK1 phosphorylation sites. Wnt treatment induces the rapid CK1γ-mediated phosphorylation of these sites within LRP6, which, in turn, promotes the recruitment of the scaffold protein Axin. Our results reveal an evolutionarily conserved mechanism that couples Wnt receptor activation to the cytoplasmic signal transduction apparatus.

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Figure 1: Identification of CK1γ as a specific modifier of LRP6.
Figure 2: CK1γ localizes at the cell membrane and associates with LRP6.
Figure 3: CK1γ phosphorylates two Ser/Thr clusters flanking a PPPSP motif in LRP6.
Figure 4: CK1γ mediates Axin recruitment and Wnt-induced phosphorylation at threonine 1479.

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Acknowledgements

We thank K. Bartscherer and M. Boutros for providing reagents and help with the Drosophila cell experiments, H. Clevers, R. Grosschedl, X. He, R. Moon, J. Nathans and R. Nusse for reagents, and H. Spring for confocal microscopy. We also thank S. Cohen and M. Boutros for comments on the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft.

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  1. Gary Davidson and Wei Wu: *These authors contributed equally to this work

Authors and Affiliations

  1. Division of Molecular Embryology, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280, D-69120, Heidelberg, Germany

    Gary Davidson, Wei Wu, Jinlong Shen, Josipa Bilic, Ursula Fenger, Peter Stannek, Andrei Glinka & Christof Niehrs

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Correspondence to Gary Davidson or Christof Niehrs.

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Competing interests

The sequence of Xenopus ck1γ2 has been deposited into GenBank (accession number DQ185136). Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure 1

Brief, schematic outline of the LRP6 modification screen that identified CK1γ as specific LRP6 kinase, characterisation of dominant negative CK1γ specificity and additional Xenopus GoF (axis duplication) as well as LoF (RT-PCR and luciferase reporter assays) experiments showing sufficiency and requirement of CK1γ in Wnt signalling. (PDF 210 kb)

Supplementary Figure 2

Vertebrate CK1 family, showing schematic comparison and amino acid sequence alignments of all members. (PDF 156 kb)

Supplementary Figure 3

Xenopus Laevis ck1γ1 and ck1γ2 mRNA expression patterns. (PDF 114 kb)

Supplementary Figure 4

Xenopus embryo LoF and GoF phenotypes showing role of CK1γ in Wnt mediated anterior-posterior patterning. (PDF 232 kb)

Supplementary Figure 5

Ser/Thr deletion analysis within ICD of FLAG-LRP6-DE(1-4)-δ87, revealing two CK1 site clusters required for Wnt signalling activity. (PDF 25 kb)

Supplementary Figure 6

Multiple sequence alignments showing evolutionary conserved nature of CK1 phosphorylation sites within LRP6 ICD and their juxtaposition with PPPS/TP priming sites. (PDF 32 kb)

Supplementary Figure 7

Characterisation of phosphor-specific T1479 and S1490 as well as phospo-independent T1479 polyclonal LRP6 peptide antibodies. (PDF 31 kb)

Supplementary Figure 8

Model for LRP6 phosphorylation by CK1γ and Axin recruitment. (PDF 118 kb)

Supplementary Table 1

List of statistics for morphological phenotypes obtained in Xenopus embryos corresponding to CK1γ GoF and LoF experiments shown in figures. (PDF 36 kb)

Supplementary Notes

This file contains the Supplementary Figure Legends and Supplementary Methods. (PDF 39 kb)

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Davidson, G., Wu, W., Shen, J. et al. Casein kinase 1 γ couples Wnt receptor activation to cytoplasmic signal transduction. Nature 438, 867–872 (2005). https://doi.org/10.1038/nature04170

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