FIG 1.

MT binding of YAP and TAZ. (A) MT-103A is defective in binding to YAP and TAZ. 293T cells were transiently transfected with vectors encoding MT, FLAG-YAP, or HA-TAZ. After ∼40 h, YAP or TAZ was immunoprecipitated with antibodies to the tag. After SDS-PAGE, Western blotting was carried out to detect YAP, TAZ, and MT. (B) MT and ST have similar affinities for YAP and TAZ. Full-length wild-type (WT) MT and ST were cotransfected in 293T cells in the presence of FLAG-YAP or HA-TAZ. YAP or TAZ was immunoprecipitated from cell extracts 40 h later. After SDS-PAGE, Western blot analysis was carried out to detect MT and ST and compare the relative ratios of MT to ST between cell extracts and YAP or TAZ complexes. WCL, whole-cell lysate. (C) The WW domains of YAP are involved in binding MT. A diagram of the YAP domain structure is shown. Full-length YAP, YAP mutated in the first WW domain (W199A and P202A), YAP mutated in the two WW domains (W199A, P202A, W258A, and P261A), and YAP with a 5-residue C-terminal deletion were each cotransfected with MT in 293T cells. After SDS-PAGE, Western blot analysis of MT complexes was carried out to detect MT and YAP.