Transcription start sites were determined by primer extension analysis. A sequence matching 7 out of 10 nucleotides of the IS box motif was identified by visual inspection of the pvdF promoter region. Putative PvdS binding sites were identified by measuring promoter activities of the pvdE and pvdF promoter deletion fragments with lacZ reporter assays. The mutation to the IS box nearest to pvdF abolished expression from the pvdF promoter and also greatly reduced expression from the pvdE promoter. In vitro transcription assays with wild type and promoters containing mutated PvdS binding sites confirmed PvdS-mediated expression. EMSAs confirmed direct regulation of pvdE-pvdF and pvdD by PvdS.
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|