Fig. 3. Interaction of NOT4 with the N-terminal region of Nos. (A) CycB (green) and Vasa (red) accumulation in embryos from wild-type (wt) and CCR4 ^KG877/Df(3R)crb-F89-4 females. (B) GST-pulldown experiments show that Hs Pum1 and Dm Pum interact in a similar fashion with three different Pop2 homologs; each GST fusion was incubated with Hs CNOT7 (lane 1), Hs CNOT8 (lane 2) and Dm Pop2 (lane 3). Note that a truncated fragment of CNOT8 generated during in vitro synthesis binds preferentially to both Pum RBDs. (C) Four different fragments of NOT4 interact with full-length Nos in yeast, as does Cup [supporting growth -His + 1 mM 3-aminotriazole (3-AT)]. At the bottom, interaction of NOT4 residues 597-1051 with various portions of Nos is summarized on the left, based on growth -His + 5 mM 3-AT, shown on the right. Inclusion of residues 1-42 of Nos is necessary for efficient expression of the C-terminal zinc finger domain (ZF) in yeast. The AD-Cup fusion provides a positive control and the AD encoded by the empty pGAD424 vector provides a negative control.