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The KLHL12–Cullin-3 ubiquitin ligase negatively regulates the Wnt–β-catenin pathway by targeting Dishevelled for degradation

Nature Cell Biology volume 8, pages 348–357 (2006)Cite this article

Abstract

Dishevelled is a conserved protein that interprets signals received by Frizzled receptors. Using a tandem-affinity purification strategy and mass spectrometry we have identified proteins associated with Dishevelled, including a Cullin-3 ubiquitin ligase complex containing the Broad Complex, Tramtrack and Bric à Brac (BTB) protein Kelch-like 12 (KLHL12). This E3 ubiquitin ligase complex is recruited to Dishevelled in a Wnt-dependent manner that promotes its poly-ubiquitination and degradation. Functional analyses demonstrate that regulation of Dishevelled by this ubiquitin ligase antagonizes the Wnt–ß-catenin pathway in cultured cells, as well as in Xenopus and zebrafish embryos. Considered with evidence that the distinct Cullin-1 based SCFβ-TrCPcomplex regulates β-catenin stability, our data on the stability of Dishevelled demonstrates that two distinct ubiquitin ligase complexes regulate the Wnt–ß-catenin pathway.

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Figure 1: Identification of the KLHL12–Cullin3 E3 ligase as a Dishevelled interacting complex.
Figure 2: Interaction of KLHL12 with Cullin-3 and Dsh.
Figure 3: KLHL12 is a negative regulator of Wnt and Dsh signalling.
Figure 4: KLHL12 recruits Dsh to Cullin-3 for protein degradation.
Figure 5: Ternary complex requirement for the ubiquitination of Dsh.
Figure 6: Analysis of KLHL12 function in zebrafish embryos.
Figure 7: Analysis of KLHL12 function in Xenopus embryos.

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Acknowledgements

We thank A.-C. Gingras for insightful technical discussions. S.A. is an associate and R.T.M. an investigator of the Howard Hughes Medical Institute. M.M. was supported by National Institutes of Health-National Center for Research Resources (NIH-NCRR) grant (P41 RR011823).

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Authors and Affiliations

  1. Howard Hughes Medical Institute, University of Washington School of Medicine, Box 357370, Seattle, 98195, WA, USA

    Stephane Angers, Chris J. Thorpe, Travis L. Biechele & Randall T. Moon

  2. Department of Pharmacology, Center for Developmental Biology, University of Washington School of Medicine, Box 357370, Seattle, 98195, WA, USA

    Stephane Angers, Chris J. Thorpe, Travis L. Biechele, Seth J. Goldenberg, Ning Zheng & Randall T. Moon

  3. Department of Genome Sciences, University of Washington School of Medicine, Box 357370, Seattle, 98195, WA, USA

    Michael J. MacCoss

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Angers, S., Thorpe, C., Biechele, T. et al. The KLHL12–Cullin-3 ubiquitin ligase negatively regulates the Wnt–β-catenin pathway by targeting Dishevelled for degradation. Nat Cell Biol 8, 348–357 (2006). https://doi.org/10.1038/ncb1381

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