Cholesterol Binding is a Determinant of Canonical Wnt Signaling Activity of Dishevelled

Dishevelled (Dvl) serves as a key scaffold protein in both canonical and non‐canonical Wnt signaling pathways. However, the molecular mechanism that determines how differently Dvl participates in these pathways remains elusive. Here, we show that Dvl is a novel cholesterol binding protein and its cholesterol binding is specifically required for the canonical Wnt signaling branch. Dvl interacts with cholesterol with high affinity and specificity via its PDZ domain. In Xenopus embryo, cholesterol binding of Dvl2 is required for the canonical Wnt‐dependent cellular and developmental processes but not for those mediated by non‐canonical Wnt/planar cell polarity (PCP) signaling. Confocal and single molecule imaging shows that cholesterol binding is essential for sustained association of Dvl2 with the Frizzled (Fz) receptor in response to canonical Wnt ligands, enabling its co‐localization with the activated Wnt receptor complex containing low‐density lipoprotein receptor‐related protein6 (LRP6). Taken together, our results suggest that cholesterol serves as a lipid signal that directs canonical Wnt pathway‐specific functions of Dvl.