FIGURE 7.

Identification of a TvCyP1-binding motif in Myb1. N-terminal deletion (A) and site-directed mutagenesis (B) were introduced into pBait-Myb1/R3(2)C to identify the TvCyP1-binding motif in Myb1 by a bacterial two-hybrid assay in which the bait plasmids were each incubated with pTRG-c21. The relative strength of the interactions as revealed by the formation of colonies in each assay is summarized in the right panel (− indicates no colony formation, + indicates <50 colonies, ++ indicates 50–100 colonies, and +++ indicates >300 colonies). In C, recombinant His-Myb1 and its derived mutant proteins His-Myb1(G106A) and His-Myb(P107A) were respectively incubated with GST and GST-TvCyP1 and immobilized on glutathione beads. The reaction products and 10% input of His-Myb1 and derived mutants were examined by Western blotting using the anti-His₆ antibody.