CAP and Nag repressor binding to the regulatory regions of the nagE-B and manX genes of Escherichia coli

Abstract

The divergent nagE-BACD operons located at 15.5 min on the Escherichia coli chromosome encode genes involved in the uptake and metabolism of N-acetylglucosamine. The start sites of the divergent transcripts are separated by 133 base-pairs (bp). A repressor protein for the regulon is encoded by the gene nagC, one of the genes of the nagBACD operon. Strains overproducing the NagC protein have been used to investigate the binding of repressor to the intergenic nagE-B regulatory region. Two binding sites have been detected, overlapping the promoters of the nagE and nagB genes. NagC binding produces a series of DNase I hypersensitive sites separated by 9 to 11 bp in the region between the two NagC binding sites, supporting a model where the NagC proteins bind co-operatively to these two sites on the DNA and interact to form a DNA loop. A strong CAP binding site exists between the two operator sites. It is located at -61.5 and -71.5 relative to the nagE and nagB transcription start sites. CAP and NagC can bind simultaneously and produce a complex more stable than the binary NagC-DNA complex. In addition NagC and CAP binding sites have been found upstream from the manXYZ operon. Although the sites exhibit a similar organization there is no evidence for formation of a DNA loop in this operon.