EMSA analysis showed that VqsR binds the qscR promoter. DNase I footprinting assay showed that VqsR binds to a region in the qscR promoter that contains an 18-bp inverted repeat motif. EMSA analysis showed that VqsR binding to the qscR promoter was abolished when this IR motif was deleted. A site-directed mutagenesis of the putative VqsR binding site further confirmed that the sequence of the inverted repeat motif is recognized by VqsR. A qscR promoter-lux fusion assay in a WT PAO1 and a vqsR mutant showed that VqsR negatively regulates qscR expression. Further, VqsR-regulated IR sequence was used to perform a genome wide search of the P. aeruginosa for additional VqsR-regulated promoters. The identified promoters were verified by performing EMSAs.
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|