Multiple sequence alignment of previously defined TodT binding sites yielded a consensus sequence for TodT binding. This sequence was used for a consensus search, which yielded potential TodT binding sites in the todX promoter. DNase I footprinting of the PtodX promoter identified that TodT protected three regions on the todX promoter. Site-directed mutagenesis of the three BOX sites combined with lacZ fusion assays determined the role of TodT as an activator of PtodX transcription. EMSA confirmed the specificity of TodT binding. ITC verified binding specificity of TodT to its target sites. Alignment of all three TodT binding sites further defined the TodT binding motif.
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|