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-beta-catenin pathway in cultured cells, as well as in Xenopus and zebrafish embryos. Considered with evidence that the distinct Cullin-1 based SCF(beta-TrCP)complex regulates beta-catenin stability, our data on the stability of Dishevelled demonstrates that two distinct ubiquitin ligase complexes regulate the Wnt-beta-catenin pathway.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
MeSH terms
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Adaptor Proteins, Signal Transducing / metabolism*
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Animals
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Blotting, Western
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Carrier Proteins / physiology*
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Cell Cycle Proteins / physiology*
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Cell Line
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Chromatography, Affinity
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Cullin Proteins / physiology*
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Dishevelled Proteins
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Embryo, Nonmammalian / metabolism
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Fluorescent Antibody Technique, Indirect
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Humans
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Intracellular Signaling Peptides and Proteins
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Kidney / cytology
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Microscopy, Fluorescence
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Phosphoproteins / metabolism*
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Signal Transduction*
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Ubiquitin
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Ubiquitin-Protein Ligases / physiology*
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Wnt Proteins / metabolism*
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Xenopus Proteins / physiology*
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Xenopus laevis / embryology
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Zebrafish / embryology
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Zebrafish Proteins / physiology*
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beta Catenin / metabolism*
Substances
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Adaptor Proteins, Signal Transducing
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CUL3 protein, human
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Carrier Proteins
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Cell Cycle Proteins
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Cullin Proteins
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Dishevelled Proteins
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Intracellular Signaling Peptides and Proteins
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KLHL12 protein, Xenopus
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KLHL12 protein, zebrafish
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Phosphoproteins
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Ubiquitin
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Wnt Proteins
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Xenopus Proteins
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Zebrafish Proteins
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beta Catenin
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Ubiquitin-Protein Ligases