Figure 1.

Ligase activation is polymerization-independent in membrane-tethered dishevelled. Shown are Western blots of His pulldowns from HEK293T cells co-transfected with His-Ub, FLAG-Dvl2 (WT, M2, PPXY mutant, or M2-PPXY mutant) and HA-tagged WT or catalytically dead (C/A) WWP2, as indicated above the panels. The left panel shows the experiment using soluble Dvl2 compared with membrane-targeted CD8-Dvl2 (right panel). WWP2 autoubiquitination is induced by WT but not PPXY mutant or polymerization-defective (M2) Dvl2; membrane-targeted Dvl2 induces WWP2 autoubiquitination and degradation even in the absence of polymerization (CD8-M2-Dvl2) (arrows). The blots are representative of three independent experiments. MW, molecular weight.