Site directed mutagenesis was used to create a strain with increased chvI function. Microarray analysis discovered that 55 genes were activated by ChvI and 4 genes are repressed by ChvI. GUS reporter gene assay was used on 7 of those genes (ropB1, SMb21188, SMb21440, SMb21491, SMc01580, SMc01855, SMc00159) and the results validated the results obtained from the microarray analysis. EMSA determined direct binding upstream of the SMc01580 gene. Direct binding was also observed upstream of the ropB1 gene and the SMb21440. DNase I footprinting localized the region bound by ChvI in the SMc01580 promoter. MEME algorithm identified an 11-bp-long motif that was present at least once in each of the ChvI-regulated genes from the microarray analysis
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 |
---|---|---|---|---|---|
AGCGTGCCTTTTTTTCGCCACAA | SMc01580, |
|
activator | not specified |