Transcription start point mapped by primer extension assay. EMSA with phosphorylated DevR in intergenic region demonstrated binding. Confirmed via DNA pulldown (immuno-precipitation). DNAse I footprinting to specify binding regions, D1, D2, D3 boxes. Mutation at all boxes decreased binding as measured by DNAse footprinting. Promoter region fused with promoterless GFP and promoter activity was detected in hypoxic conditions through GFP reporter assay. Mutations at all three sites decreased promoter activity as measured by GFP reporter assay.
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|