Fig. 1.

Autoinhibition of Daam1 is mediated by interactions between its amino-terminal and carboxyl-terminal regions. (A) GST pull-down assays reveal that T-Daam binds to the PDZ domain of Dvl and to N-Daam. Precipitated T-Daam was detected with Western blotting, and input of GST proteins was visualized by using Coomassie staining. (B–D) Coimmunoprecipitation assays. Plasmids encoding tagged Daam1 fragments were cotransfected into HEK293T cells, and cell lysates were immunoprecipitated (IP) with indicated Abs. Precipitates were then immunoblotted with indicated Abs. (B) N-Daam interacts with T-Daam. (C) N-Daam interacts with C-Daam, but this interaction is reduced with C-DaamΔDAD, which lacks the DAD. (D) Mutations within the DAD in the context of C-Daam reduces interaction with N-Daam.