Figure 3
From: WW domain-mediated interaction with Wbp2 is important for the oncogenic property of TAZ
Interaction of Wbp2 with TAZ depends on the PPXY motifs-containing C-terminal region of Wbp2. (a) The schematic diagram shows the N-terminal, C-terminal regions of Wbp2 and the positions of three PPXY motifs. (b) The C-terminal region of Wbp2 is important for interaction with TAZ. Whole-cell lysates from 293 cells transiently expressing Flag-TAZ together with GFP-Wbp2, GFP-Wbp2-N-terminal or GFP-Wbp2-C-terminal regions were immunoprecipitated with anti-Flag, and the immunoprecipitates were analyzed by immunoblot to detect coimmunoprecipitated GFP-Wbp2 and its mutants. The blots were stripped and re-probed with anti-Flag-HRP to detect immunoprecipitated Flag-TAZ. (c) PPXY motifs (aa197-200) of Wbp2 are important for interaction with TAZ. In all, 293 cells expressing Flag-TAZ and various mutants of HA-Wbp2 were processed for immunoprecipitation. The immunoprecipitates were analyzed by immunoblot to detect coimmunoprecipitated Wbp2 and its PPXY mutants. Mutation of the second PPXY motif significantly reduced the ability of Wbp2 to interact with TAZ. Lane 1 is negative control for immunoprecipitation with 293 cells expressing vector and wild-type Wbp2 only.
