EMSA confirmed that BldD bound specifically to the promoter of the erythromycin biosynthetic gene cluster. DNase I footprinting localized BldD binding region in the eryBVI promoter. The authors identified a putative BldD binding site in this region based on the S.coelicolor BldD consensus, AGTgA-(n)m-TCACc. Deletion of bldD resulted in in the inability of the strain to form aerial mycelium and to sporulate on three different media. Complementation of the bldD deletion with a single copy of bldD restored the WT phenotype. Furthermore, bldD mutant produced 7-fold less erythromycin than the WT strain. Altogether these data allowed the authors to confirm BldD-mediated activation of the ery genes.
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|