CcpA-mediated regulation was confirmed by degU-lacZ reporter assays. The induction of degU by glucose was abolished by disruption of ccpA. EMSA analysis confirmed direct binding of CcpA to the degU promoter region. DNase I footprinting confirmed that CcpA binds to cre sequence in the degU promoter. Site-directed mutagenesis of the CcpA binding site resulted in significant reduction in binding. The functionality of the CcpA binding sequence was confirmed in vivo by lacZ reporter assays in conjunction with site-directed mutagenesis.
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|