CsoR binding sites in the genome sequence of Streptomyces coelicolor A3(2) were predicted using a comparative genomics approach. Three target sequences were selected for in vitro analyses: AAATACCCCTGGTGGGTATAT, located -42 bp upstream TLS of csoR TTATACCCCCTAGGGGTAAGG, located -25 np upstream TLS of SCO2730 GGGTACCCCCTAGGGGTATAC, located -83 bp upstream TLS of SCO1045 Binding of S. lividans CsoR to these proteins was validated by EMSA using w-t and mutated protein. RNA-seq was performed and all three genes were shown to be induced under copper induction and on a csoR- background. Expression of csoR, not apparent in RNA-seq was confirmed to be CsoR/copper mediated by promoter probing with a undecylprodigiosin (redD) 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 |
---|---|---|---|---|---|
AAATACCCCTGGTGGGTATAT | SCO4136, |
|
repressor | tetramer | |
TTATACCCCCTAGGGGTAAGG | SCO2730, SCO2731, |
|
repressor | tetramer | |
GGGTACCCCCTAGGGGTATAC | SCO1045, SCO1046, |
|
repressor | tetramer |