Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin

BACKGROUND: Dishevelled-3 (Dvl3) is a multivalent scaffold essential to cell signaling in development. Dsh/Dvls enable a myriad of protein-protein interactions in Wnt signaling. In the canonical Wnt/β-catenin pathway specifically, Dvl3 polymerizes to form dynamic protein aggregates, so-called “signa...

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Autores principales: Yokoyama, Noriko, Markova, Nelli G, Wang, Hsien-yu, Malbon, Craig C
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3542119/
https://www.ncbi.nlm.nih.gov/pubmed/22748080
http://dx.doi.org/10.1186/1750-2187-7-8
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author Yokoyama, Noriko
Markova, Nelli G
Wang, Hsien-yu
Malbon, Craig C
author_facet Yokoyama, Noriko
Markova, Nelli G
Wang, Hsien-yu
Malbon, Craig C
author_sort Yokoyama, Noriko
collection PubMed
description BACKGROUND: Dishevelled-3 (Dvl3) is a multivalent scaffold essential to cell signaling in development. Dsh/Dvls enable a myriad of protein-protein interactions in Wnt signaling. In the canonical Wnt/β-catenin pathway specifically, Dvl3 polymerizes to form dynamic protein aggregates, so-called “signalsomes”, which propagate signals from the Wnt receptor Frizzled to downstream elements. RESULTS: Very large Dvl3-based supermolecular complexes form in response to Wnt3a. These complexes are identified by steric-exclusion chromatography, affinity pull-downs, proteomics, and fluorescence correlation microscopy (fcs). In the current work, the roles of Dvl3 phosphorylation and of Axin in the assembly of Dvl3-based supermolecular complexes in response to Wnt3a are probed in totipotent mouse F9 teratocarcinoma cells. Point mutations of phosphorylation sites of Dvl3 which interfere with Lef/Tcf-sensitive transcriptional activation by Wnt3a are shown to interfere more proximally with the assembly of Dvl3-based supermolecular complexes. Axin, a Dvl-interacting protein, plays a central role in organizing the beta-catenin destruction complex. The assembly of Dvl3-based supermolecular complexes is blocked either by depletion of Axin or by mutation of Axin sites necessary for polymerization in response to Wnt3a. CONCLUSION: These data demonstrate that Wnt3a activation of the canonical pathway requires specific phosphorylation events as well as Axin to assemble very large, Dvl3-based supermolecular complexes; these complexes are a prerequisite to activation of Lef/Tcf-sensitive transcription.
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spelling pubmed-35421192013-01-11 Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin Yokoyama, Noriko Markova, Nelli G Wang, Hsien-yu Malbon, Craig C J Mol Signal Research Article BACKGROUND: Dishevelled-3 (Dvl3) is a multivalent scaffold essential to cell signaling in development. Dsh/Dvls enable a myriad of protein-protein interactions in Wnt signaling. In the canonical Wnt/β-catenin pathway specifically, Dvl3 polymerizes to form dynamic protein aggregates, so-called “signalsomes”, which propagate signals from the Wnt receptor Frizzled to downstream elements. RESULTS: Very large Dvl3-based supermolecular complexes form in response to Wnt3a. These complexes are identified by steric-exclusion chromatography, affinity pull-downs, proteomics, and fluorescence correlation microscopy (fcs). In the current work, the roles of Dvl3 phosphorylation and of Axin in the assembly of Dvl3-based supermolecular complexes in response to Wnt3a are probed in totipotent mouse F9 teratocarcinoma cells. Point mutations of phosphorylation sites of Dvl3 which interfere with Lef/Tcf-sensitive transcriptional activation by Wnt3a are shown to interfere more proximally with the assembly of Dvl3-based supermolecular complexes. Axin, a Dvl-interacting protein, plays a central role in organizing the beta-catenin destruction complex. The assembly of Dvl3-based supermolecular complexes is blocked either by depletion of Axin or by mutation of Axin sites necessary for polymerization in response to Wnt3a. CONCLUSION: These data demonstrate that Wnt3a activation of the canonical pathway requires specific phosphorylation events as well as Axin to assemble very large, Dvl3-based supermolecular complexes; these complexes are a prerequisite to activation of Lef/Tcf-sensitive transcription. BioMed Central 2012-06-29 /pmc/articles/PMC3542119/ /pubmed/22748080 http://dx.doi.org/10.1186/1750-2187-7-8 Text en Copyright ©2012 Yokoyama et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Yokoyama, Noriko
Markova, Nelli G
Wang, Hsien-yu
Malbon, Craig C
Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin
title Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin
title_full Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin
title_fullStr Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin
title_full_unstemmed Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin
title_short Assembly of Dishevelled 3-based supermolecular complexes via phosphorylation and Axin
title_sort assembly of dishevelled 3-based supermolecular complexes via phosphorylation and axin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3542119/
https://www.ncbi.nlm.nih.gov/pubmed/22748080
http://dx.doi.org/10.1186/1750-2187-7-8
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