Transcriptional control of the cysG gene of Escherichia coli K-12 during aerobic and anaerobic growth.
The 74-min region of the Escherichia coli chromosome includes five open reading frames of known sequence. The first and last of these genes, nirB and cysG, are transcribed in the same direction and both are essential for NADH-dependent nitrite reductase activity. The functions of the other genes, nirD, nirE and nirC, which are located between nirB and cysG, are unknown. The nirB gene is transcribed from a promoter which is anaerobically induced, expression being dependent on the transcription activator protein, Fnr. Here we show that the nirD, nirE, nirC and cysG genes are also expressed from the nirB promoter. After subcloning cysG, a second promoter was located less than 100 bases upstream of cysG. Two groups of transcription start points separated by 40 bases were detected in this region by S1 mapping. Rates of transcription from the isolated cysG promoter were the same during aerobic growth and anaerobic growth in the presence or absence of nitrite. However, when the nirB gene and its promoter were cloned back upstream from the cysG promoter, the rate of transcription was higher during anaerobic growth than during aerobic growth and was further induced by nitrite. These increases were totally dependent on a functional fnr gene and were shown by S1 mapping experiments to be due to transcriptional read-through from the Fnr-dependent nirB promoter. No promoter activity was associated with DNA fragments between the BamHI site located within the N-terminal coding region of the nirB gene and the cysG promoter located at the C-terminus of nirC.
Peakman
T
T
School of Biochemistry, University of Birmingham, England.
Busby
S
S
Cole
J
J
D000332:Aerobiosis; D000595:Amino Acid Sequence; D000693:Anaerobiosis; D002876:Chromosomes, Bacterial / Q000032:analysis*; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000235:genetics; D010449:Peptide Mapping; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis*; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D012333:RNA, Messenger; D009565:Nitrate Reductases; D050901:Nitrate Reductase