Proteomic Analysis of Wnt-Dependent Dishevelled-Based Supermolecular Complexes

Yokoyama, Noriko

doi:10.5772/30271

Cited by 2 publications

(4 citation statements)

References 85 publications

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“…PI4K, together with PIP5K, rapidly produces PIP 2 after Wnt stimulation and enhances the phosphorylation of LRP5/6 and oligomerization of Dishevelled and LRP5/6 (Pan et al, 2008). Proteomic analysis also showed that Dishevelled interacts with PI4K, Rho, and even G α13 , after Wnt stimulation (Yokoyama, 2012). In addition, trafficking of PI4K and PIP5K to the plasma membrane is regulated by Rho GTPase (Weernink et al, 2004;Yang et al, 2004).…”

Section: Resultsmentioning

confidence: 96%

“…LGR5, first identified as a member of the glycoprotein hormone receptor family (Hsu et al, 1998), had been an orphan receptor until several groups showed that R-spondins (RSPOs) were able to bind and activate LGR5, enhancing canonical Wnt-β-catenin signaling together with LRP 5/6 and Frizzled (Carmon et al, 2011;2012;de Lau et al, 2011;Glinka et al, 2011;Ruffner et al, 2012). LGR5 is expressed and plays an important role in various adult tissue stem cells in, for example, small intestine, hair follicles, and stomach, through regulation of Wnt signaling (Barker and Clevers, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Leucine-Rich Repeat-Containing G-Protein Coupled Receptor 5/GPR49 Activates G12/13-Rho GTPase Pathway

Kwon

Park

Kim

et al. 2013

Molecules and Cells

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Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5/GPR49) is highly expressed in adult stem cells of various tissues, such as intestine, hair follicles, and stomach.LGR5 is also overexpressed in some colon and ovarian tumors. Recent reports show that R-spondin (RSPO) family ligands bind to and activate LGR5, enhancing canonical Wnt signaling via the interaction with LRP5/6 and Frizzled. The identity of heterotrimeric G-proteins coupled to LGR5, however, remains unclear. Here, we show that Rho GTPase is a downstream target of LGR5. Overexpression of LGR5 induced SRF-RE luciferase activity, a reporter of Rho signaling. RSPOs, ligands for LGR4, LGR5, and LGR6, however, did not induce SRF-RE reporter activity in the presence of LGR5. Consistently, LGR5-induced activity of the SRF-RE reporter was inhibited by Rho inhibitor C3 transferase and RhoA N19 mutant, and knockdown of Gα 12/13 genes blocked the reporter activity induced by LGR5. In addition, focal adhesion kinase, NF-κB and c-fos, targets of Rho GTPase, were shown to be regulated by LGR5. Here, we have demonstrated, for the first time, that LGR5 is coupled to the Rho pathway through G 12/13 signaling.

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Section: Resultsmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Leucine-Rich Repeat-Containing G-Protein Coupled Receptor 5/GPR49 Activates G12/13-Rho GTPase Pathway

Kwon

Park

Kim

et al. 2013

Molecules and Cells

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Section: Discussionmentioning

confidence: 99%

“…Proteomic analyses of Dvl3-based supermolecular complexes separated by SEC identify CK1α, CK1γ, CK2α and β, LRP6, PI4 kinase and Src [39]. The proteomic data derived from isolated supermolecular complexes reinforces and extends the functional significance of Dvl3-based supermolecular complexes [39]. The upper limit of Dvl3-based supermolecular complexes has been amended recently to M r = 35 MDa, based upon estimates recalibrated from fluorescence correlation microscopy in live cells [40].…”

Section: Discussionmentioning

confidence: 99%

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

Yokoyama

Markova²,

Wang

et al. 2012

JMS

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BackgroundDishevelled-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.ResultsVery 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.ConclusionThese 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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Proteomic Analysis of Wnt-Dependent Dishevelled-Based Supermolecular Complexes

Cited by 2 publications

References 85 publications

Leucine-Rich Repeat-Containing G-Protein Coupled Receptor 5/GPR49 Activates G12/13-Rho GTPase Pathway

Leucine-Rich Repeat-Containing G-Protein Coupled Receptor 5/GPR49 Activates G12/13-Rho GTPase Pathway

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

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