Interaction of Axin and Dvl-2 proteins regulates Dvl-2-stimulated TCF-dependent transcription

Smalley, Matthew J.; Sara, E. A.; Paterson, Hugh; Naylor, Stuart; Cook, David P.; Jayatilake, Hiran; Fryer, Lee G.D.; Hutchinson, Lisa; Fry, Michael; Dale, Trevor

doi:10.1093/emboj/18.10.2823

Cited by 228 publications

(240 citation statements)

References 46 publications

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“…Using a commercially available polyclonal axin antibody, we observed endogenous axin in distinct puncta in the cytoplasm of Madin-Darby canine kidney (MDCK) cells (Figures 1b and c). Both endogenous and recombinant axin puncta were evenly distributed throughout the cytoplasm (Figures 1b-e), consistent with previous reports for endogenous axin in HEK293 cells (Levina et al, 2004;Wiechens et al, 2004) and recombinant axin (Fagotto et al, 1999;Smalley et al, 1999;Schwarz-Romond et al, 2005, 2007b. Axin-RFP was localized to cytoplasmic puncta when expressed in both MDCK ( Figure 1e) and HEK293 T cells (Supplementary Figures S1, S2).…”

Section: Resultssupporting

confidence: 90%

“…The ability of axin to form puncta has been reported previously (Fagotto et al, 1999;Smalley et al, 1999;Schwarz-Romond et al, 2005), and recent studies show that puncta formation is a dynamic process mediated by protein oligomerization or polymerization (SchwarzRomond et al, 2007a, b). Our data provide evidence for the existence of endogenous axin puncta, consistent with the detection of small punctate structures or spots with axin antibodies in 293 cells (Levina et al, 2004;Wiechens et al, 2004).…”

Section: Discussionmentioning

confidence: 76%

“…Endogenous axin has been detected in small, punctate cytoplasmic structures that redistribute to large aggregates close to the plasma membrane following treatment with LiCl (which inhibits GSK3-b, mimicking Wnt activation) (Levina et al, 2004;Wiechens et al, 2004). Ectopically expressed recombinant axin is localized in cytoplasmic vesicles or puncta (Fagotto et al, 1999;Smalley et al, 1999;Schwarz-Romond et al, 2005). The C-terminal DIX domain of axin is known to mediate self-association, which is thought to be important for the regulation of b-catenin (Hsu et al, 1999;Kishida et al, 1999;Sakanaka and Williams, 1999) and has recently been shown to associate into polymers that are required for puncta formation (Schwarz-Romond et al, 2007a, b).…”

Section: Introductionmentioning

confidence: 99%

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Recruitment of adenomatous polyposis coli and β-catenin to axin-puncta

et al. 2008

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The adenomatous polyposis coli (APC) tumour suppressor is a multifunctional protein involved in the regulation of Wnt signalling and cytoskeletal dynamics. Little is known about how APC controls these disparate functions. In this study, we have used APC-and axin-fluorescent fusion proteins to examine the interactions between these proteins and show that the functionally distinct populations of APC are also spatially separate. Axin-RFP forms cytoplasmic punctate structures, similar to endogenous axin puncta. Axin-RFP recruits b-catenin destruction complex proteins, including APC, b-catenin, glycogen synthase kinase-3-b (GSK3-b) and casein kinase-1-a (CK1-a). Recruitment into axin-RFP puncta sequesters APC from clusters at cell extensions and this prevents its microtubule-associated functions. The interaction between APC-GFP and axin-RFP within the cytoplasmic puncta is direct and dramatically alters the dynamic properties of APC-GFP. However, recruitment of APC to axin puncta is not absolutely required for b-catenin degradation. Instead, formation of axin puncta, mediated by the DIX domain, is required for b-catenin degradation. An axinDDIX mutant did not form puncta, but still mediated recruitment of destruction complex proteins and phosphorylation of b-catenin. We conclude that there are distinct pools of APC and that the formation of axin puncta, rather than the axin/APC complex, is essential for b-catenin destruction.

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Section: Resultssupporting

confidence: 90%

Section: Discussionmentioning

confidence: 76%

Section: Introductionmentioning

confidence: 99%

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Recruitment of adenomatous polyposis coli and β-catenin to axin-puncta

et al. 2008

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“…Functional interaction between WWOX and Wnt pathway N Bouteille et al WWOX does not inhibit the interaction of Dvl-2 with the b-catenin-degradation complex In the b-catenin-degradation complex, Dvl-2 binds directly to Axin. This interaction seems to be crucial for Dvl-2 to inhibit glycogen synthase kinase 3b-dependent degradation of b-catenin (Kishida et al, 1999Smalley et al, 1999;Kadoya et al, 2000;Hino et al, 2001). WWOX would therefore inhibit the function of Dvl-2 in the Wnt/b-catenin pathway by disrupting Dvl-2-Axin association.…”

Section: Binding Domainsmentioning

confidence: 99%

Inhibition of the Wnt/β-catenin pathway by the WWOX tumor suppressor protein

et al. 2009

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The WWOX gene encodes a candidate tumor suppressor protein (WWOX) implicated in a variety of human diseases such as cancer. To better understand the molecular mechanisms of WWOX action, we investigated novel partners of this protein. Using the two-hybrid system and a coimmunoprecipitation assay, we observed a physical association between WWOX and the Dishevelled protein (Dvl) family signaling elements involved in the Wnt/b-catenin pathway. We found that enforced WWOX expression inhibited, and inhibition of endogenous WWOX expression stimulated the transcriptional activity of the Wnt/b-catenin pathway. Inhibition of endogenous WWOX expression also enhanced the effect of Wnt-3a on b-catenin stability. Moreover, we observed the sequestration of Dvl-2 wild type and Dvl-2NESm, a mutated form of Dvl-2 predominantly localized in the nucleus, in the cytoplasm compartment by WWOX. Our results indicate that WWOX is a novel inhibitor of the Wnt/b-catenin pathway. WWOX would act, at least in part, by preventing the nuclear import of the Dvl proteins.

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“…The results of Dsh or Dvl overexpression can also mimic the effects of Wg in cultured cells 9,10 , in the eye 11 , and in the heart 12 . In addition, recent results have provided exciting insight into the mechanisms by which activated Dsh and Dvl proteins lead to the suppression of GSK-3β activity [13][14][15][16][17][18] .…”

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confidence: 99%

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et al. 2000

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The DEP domain of Dishevelled (Dvl) proteins transduces signals to effector proteins downstream of Dvl in the Wnt pathway. Here we report that DEP-containing mutants inhibit Wnt-induced, but not Dvl-induced, activation of the transcription factor Lef-1. This inhibitory effect is weakened by a K434M mutation. Nuclear magnetic resonance spectroscopy revealed that the DEP domain of mouse Dvl1 comprises a three-helix bundle, a β-hairpin 'arm' and two short β-strands at the Cterminal region. Lys 434 is located at the tip of the β-hairpin 'arm'. Based on our findings, we conclude that DEP interacts with regulators upstream of Dvl via a strong electric dipole on the molecule's surface created by Lys 434, Asp 445 and Asp 448; the electric dipole and the putative membrane binding site are at two different locations.The Wnt signaling pathway is a key regulatory pathway for cellular development and growth. Wnt signaling has been extensively studied in Drosophila, Caenorhabditis elegans, Xenopus, and mammalian systems [1][2][3][4][5] . Through genetic studies, a working model of the Wnt signaling pathway has been established. Secreted Wnt proteins bind to Frizzled transmembrane receptors. The receptor-ligand complex activates the Dishevelled (Dsh and Dvl) proteins, which suppress the activity of glycogen synthase kinase-3β (GSK-3β). If its activity is not suppressed, GSK-3β binds to adenomatous polyposis coli (APC) protein and axin and then phosphorylates specific Ser and Thr residues at the N-terminus of β-catenin. In turn, the ubiquitin-proteasome pathway rapidly degrades hyperphosphorylated β-catenin. Wnt activation prevents this process by promoting the cytosolic accumulation and subsequent translocation of β-catenin into the nucleus. Once in the nucleus, β-catenin binds to the transcription factors T-cell factor (Tcf) or lymphoid enhancer factor (Lef) and acts as a transcriptional coactivator 5 . The result is increased expression of Tcf or Lef regulated target genes, such as Myc 6 and the gene encoding cyclin D 7 .© 2000 Nature America Inc. Correspondence should be addressed to: J.Z. Jie.Zheng@stjude.org. HHS Public Access DEP domain interacts with upstream regulators of DvlWe and others have demonstrated that Wnt-1 or Dvl1, when expressed in NIH3T3 cells, can activate the transcription regulator 16,28,29). This activation depends on β-catenin and is suppressed by GSK-3β and axin. To characterize the role of the mDvl1 DEP domain in Wnt signaling, we examined the effect of the Dvl1 N-terminal truncation mutant, DvlC1, on Wnt-induced activation of the transcription factor Lef-1. In Lef dependent reporter gene assays, DvlC1 inhibited Wnt-1 induced Lef-1 activation but not Dvl1 induced Lef-1 activation (Fig. 2). To further characterize the specific sequences that inhibit the Wnt-induced activation, we generated two additional mutants of mDvl1: DvlDEP, which contains only the DEP domain; and DvlC2, which encompasses the sequence downstream of the DEP domain. When expressed in NIH3T3 cells, DvlDEP, like DvlC1, bloc...

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Interaction of Axin and Dvl-2 proteins regulates Dvl-2-stimulated TCF-dependent transcription

Cited by 228 publications

References 46 publications

Recruitment of adenomatous polyposis coli and β-catenin to axin-puncta

Recruitment of adenomatous polyposis coli and β-catenin to axin-puncta

Inhibition of the Wnt/β-catenin pathway by the WWOX tumor suppressor protein

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