Nickel regulation of H. mustelae ureA and ureA2 transcription start sites was quantified by Western blot and quantitative reverse transcriptase PCR. Putative NikR binding sites were found in the ureA and ureA2 promoters using a modified consensus sequence from H. pylori. EMSA showed NikR binding to both sites only in the presence of nickel, and incubation with the ureA2 promoter resulted in two bands, suggesting multiple NikR binding sites. The putative binding site in the ureA2 promoter was mutated to a C stretch, and EMSA indicated binding in the wild type but not the mutant. A new motif was proposed using the H. pylori high-affinity sites and these H. mustelae NikR operators. This motif was used to identify a potential NikR-regulated gene in H. mustelae, hm0418-1. Western blot confirmed its expression to be nickel-regulated.
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|
||not specified||not specified|