Fig. 10

OsWRKY80 binds to OsWRKY4 promoter in yeast. a Distribution of W-box and W-box like elements in the OsWRKY4 promoter, 1.5 Kb upstream from the translational start site. Black triangles indicate W-box or W-box like elements. A 79-nucleotide-long promoter fragment, which contains four W-box or W-box like elements, was indicated, and used for DNA-binding test in yeast. The W-box or W-box like elements are underlined and in black. Bases in red were indicated as the mutant sites. b Yeast cells were co-transformed with a bait vector, containing a promoter fragment in (a) fused to a HIS2 reporter gene, and a prey vector, containing OsWRKY80 fused to a GAL4 activation domain. Left, yeast cells carrying different constructs were grown for 2 d at 30°C in SD-Leu-Trp agar medium. Middle, schematic distribution of yeast cells carrying different vectors; 1, OsWRKY80 + PWRKY4; 2, OsWRKY80 + mPWRKY4; 3, PWRKY4 only. Right panel, positive interactions between OsWRKY80 and target DNA fragments were verified in SD-Leu-Trp-His agar medium with 30 mM 3-amino-1, 2, 4-triazole (3-AT) for suppression of background growth