Regulation of ndh expression in Escherichia coli by Fis.
The Escherichia coli ndh gene encodes NADH dehydrogenase II, a primary dehydrogenase used during aerobic and nitrate respiration. The anaerobic transcription factor FNR represses ndh expression by binding at two sites centred at -94.5 and -50.5. In vivo transcription studies using promoter fusions with 5' deletions confirmed that both FNR sites are required for maximum repression under anaerobic conditions. The histone-like protein Fis binds to three sites [centred at -123 (Fis I), -72, (Fis II) and +51 (Fis III)] in the ndh promoter. Using ndh : : lacZ promoter fusions carrying 5' deletions, or replacement mutations it is shown that Fis III is a repressing site and that Fis I and II are activating sites, with the greatest contribution from Fis II. Deletion of the C-terminal domain of the RNA polymerase alpha-subunit abolished Fis-mediated activation of ndh expression, suggesting that ndh has a Class I Fis-activated promoter. In accordance with the established pattern of Fis synthesis, ndh transcription was greatest during exponential growth. Thus, it is suggested that Fis enhances ndh expression during periods of rapid growth, by acting as a Class I activator, and that the binding of tandem FNR dimers represses ndh expression by preventing interaction of the RNA polymerase alpha-subunit with DNA and Fis.
Jackson
Laura
L
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
Blake
Timo
T
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
Green
Jeffrey
J
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D035021:Factor For Inversion Stimulation Protein / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D016296:Mutagenesis; D016297:Mutagenesis, Site-Directed; D009245:NADH Dehydrogenase / Q000235:genetics*; D010957:Plasmids; D017384:Sequence Deletion
D035021:Factor For Inversion Stimulation Protein; C088908:NADH dehydrogenase II; D009245:NADH Dehydrogenase