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Nature Cell Biology 8, 348–357 (1 April 2006) | doi:10.1038/ncb1381

The KLHL12|[ndash]|Cullin-3 ubiquitin ligase negatively regulates the Wnt|[ndash]||[beta]|-catenin pathway by targeting Dishevelled for degradation

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

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 |[agrave]| 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–|[szlig]|-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–|[szlig]|-catenin pathway.