Figure 6.

Determination of the Core DNA Binding Motif Using GAL4AD-ANAC055 Fusion.

Thirty-nine constructs were made to define the core motif of NACRS. Each base substitution construct was initially fused to the lacZ gene and integrated into the YM4271 chromosome. Yeast recombinant strains were then transformed with pAD-GAL4-ANAC055 or pAD-WT plasmids. β-Gal, β-galactosidase; Mu, Miller units.

(A) Results of liquid culture assays of nine constructs designed to facilitate an investigation of the first three nucleotides (red letters).

(B) Results of liquid culture assays of 30 constructs designed to facilitate an investigation of the next 10 nucleotides (red letters). The criterion of four units' difference, which is approximately one-third of the units' difference in the wild type, was used for analysis of the differences of β-galactosidase activities given by pAD-GAL4-ANAC055 and pAD-WT in each mutated construct. Nucleotide changes in the NACRS that had an adverse effect on the binding activity of ANAC055 protein are indicated by small letters.