Par-1 Regulates Tissue Growth by Influencing Hippo Phosphorylation Status and Hippo-Salvador Association
Figure 6
Par-1 disrupts the association of the Hpo-Sav complex in a kinase-dependent manner.
(A) Par-1 could destabilize Hpo-induced accumulation of Sav in a kinase-dependent manner. S2 cells were transfected with the indicated constructs followed by a Western blot analysis. CFP served as a loading control. (B) Par-1 inhibits the phosphorylation of Sav induced by Hpo. The mobility shift assay was employed. The loading volume was adjusted according to the total Sav protein level. (C) Par-1 disrupts the interaction of the Hpo-Sav complex. S2 cells were transfected with the indicated constructs followed by immunoprecipitation to test whether the interaction between Sav and endogenous Hpo was affected by Par-1. Note that less Sav interacted with Hpo in the presence of Par-1. (D–D″) Par-1 RNAi is incapable of inhibiting sav mutant-induced adult eye overgrowth. The clones were generated using the MARCM system. The genotypes are as following: eyflp, ubi-Gal4, UAS-GFP; FRT82B/FRT82BGal80 (D), eyflp, ubi-Gal4, UAS-GFP; FRT82B SavSH13/FRT82B Gal80 (D′), and eyflp, ubi-Gal4, UAS-GFP; Par-1-RNAi; FRT82B SavSH13/FRT82B Gal80 (D″). (E) The proposed mechanism of Par-1 regulation of the Hpo pathway (see text for further detail).