qRT-PCR confirmed that the atzSTUVW operon genes were co-transcribed. Beta-galactosidase assays with atzR deletion strains demonstrated the repressor activity of AtzR. Visual inspection of the atzT promoter region yielded a sequence identical to the AtzR binding site for atzDEF. Site-directed mutagenesis combined with beta-galactosidase assays identified that the integrity of this region is necessary for AtzR-dependent repression. This region was then validated with EMSA to demonstrate specific binding. DNase I footprinting localized the AtzR binding site.
Regulated genes for each binding site are displayed below. Gene regulation diagrams show binding sites, positively-regulated genes, negatively-regulated genes, both positively and negatively regulated genes, genes with unspecified type of regulation. For each indvidual site, experimental techniques used to determine the site are also given.
|Site sequence||Regulated genes||Gene diagram||Experimental techniques||TF function||TF type|