Journal: Science Advances

Article Title: A Wnt-induced conformational phospho-switch in DVL3 controls association with Frizzled receptors and Wnt/β-catenin signaling

doi: 10.1126/sciadv.aed8899

Figure Lengend Snippet: ( A ) Schematic representation of the proximity-labeling experimental workflow. Expression of WT and phospho-switch mutant proteins fused to a promiscuous biotin ligase (TurboID) was induced by doxycycline. Biotinylated proteins were identified by LC-MS/MS. Created in BioRender. Gybel, T. (2026) https://BioRender.com/pyxzao0 . ( B ) Volcano plots comparing proteins enriched (right) or depleted (left) in the interactomes of phospho-switch mutants in comparison to WT. ( C ) The potency of each interaction was calculated as the Euclidean distance from the origin of the volcano plots comparing the interactome of phospho-switch mutants with WT. The corresponding values were plotted as a function of the net charge (at pH 7.2) proximal to the DEP domain and fitted using a sigmoidal function to determine the charge threshold for each FZD. Axin interaction with DVL3 did not show significant difference between the phospho-switch mutants and WT. ( D ) FZD-DEP interface, pIDR2-DEP interface, and the two overlapping epitopes mapped onto the FZD-DEP structure (PDB ID: 8WMA). ( E ) Mechanistic model of DVL phospho-switch in Wnt/β-catenin signaling. Wnt stimulus results in FZD clustering, stabilization of DVL at the membrane, and more efficient CK1δ/ε-mediated DVL phosphorylation that leads to the accumulation of negative charge proximal to the DEP domain. Charge-mediated DVL intramolecular interaction competes with FZD binding, and when the charge threshold is reached, DVL dissociates from FZD (step 1). This event must be accompanied by the activation of LRP5/6 coreceptors (step 2), to propagate Wnt signal to downstream components. Created in BioRender. Gybel, T. (2026) https://BioRender.com/tremyul .

Article Snippet: The expression of DVL was induced by doxycycline (1 μg/ml; HY-N0565B, MedChemExpress) treatment for 24 hours and supplemented with 50 μM biotin (SC204706A, Santa Cruz Biotechnology) afterward.

Techniques: Labeling, Expressing, Mutagenesis, Liquid Chromatography with Mass Spectroscopy, Comparison, Membrane, Phospho-proteomics, Binding Assay, Activation Assay