Primer extension assay performed with the wild-type and putR mutant strains showed that putAP expression was decreased in the putR mutant. EMSA showed that PutR binds to the putAP promoter. DNase I footprinting assay revealed that PutR protected two regions in the putAP promoter which extended from -97 to -80 and from -139 to -123. put–luxAB transcriptional fusions with a wild-type PutR binding site and with a promoter in which the binding site was deleted confirmed the PutR activated the expression of the putAP operon.
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|