Escherichia coli K-12 genes essential for the synthesis of c-type cytochromes and a third nitrate reductase located in the periplasm

• Published in: Molecular microbiology Vol. 19; no. 3; p. 467
• Main Authors: Grove, J, Tanapongpipat, S, Thomas, G, Griffiths, L, Crooke, H, Cole, J
• Format: Journal Article
• Language: English
• Published: England 01.02.1996
• Subjects: Amino Acid Sequence; Cloning, Molecular; Cytochrome c Group - biosynthesis; Escherichia coli - enzymology; Escherichia coli - genetics; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genetic Complementation Test; Molecular Sequence Data; Mutagenesis, Insertional; Nitrate Reductase; Nitrate Reductases - biosynthesis; Nitrate Reductases - genetics; Nitrates - metabolism; Nitrates - pharmacology; Nitrites - metabolism; Nitrites - pharmacology; Operon; Oxidation-Reduction; Plasmids - chemistry; Plasmids - genetics; Promoter Regions, Genetic - genetics; Recombinant Fusion Proteins; Terminology as Topic
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1046/j.1365-2958.1996.383914.x

The 'aeg46.5' operon was originally detected as an 'anaerobically expressed gene' located at minute 46.5 on the Escherichia coli linkage map. Subsequent results from the E. coli Genome Sequencing Project revealed that the 'aeg46.5' promoter was located in the centisome 49 (minute 47) region. Downstream from this promoter are 15 genes, seven of which are predicted to encode a periplasmic nitrate reductase and eight encode proteins homologous to proteins essential for cytochrome c assembly in other bacteria. All of these genes, together with the 'aeg46.5' promoter, have been subcloned on a 20kb EcoRI fragment from Kohara phage 19D1. Evidence is presented that, as predicted, the region includes structural genes for two c-type cytochromes of mass 16kDa and 24 kDa, which are transcribed from the previously described 'aeg46.5' promoter, and that the first seven genes encode a functional nitrate reductase. We, therefore, propose that they should be designated nap (nitrate reductase in the periplasm) genes. Plasmids encoding the entire 20kb region, or only the downstream eight genes, complemented five mutations resulting in total absence of all five known c-type cytochromes in E coli, providing biochemical evidence that these are ccm (for cytochrome c maturation) genes. The ccm region was transcribed both from the FNR-dependent, NarL- and NarP-regulated nap promoter (formerly the 'aeg46.5' promoter) and from constitutive or weakly regulated promoters apparently located within the downstream nap and ccm genes.