The function of the upstream region of the sigma 54-dependent Klebsiella pneumoniae nifL promoter is sensitive to DNA supercoiling.
The positive control protein NTRC activates transcription from the sigma 54-dependent nifL and glnAp2 promoters of Klebsiella pneumoniae by binding to upstream enhancer-like sequences and contacting downstream bound sigma 54-RNA polymerase via looping of the intervening DNA. In contrast to the glnAp2 promoter, the activity of the nifL promoter is very sensitive to changes in DNA supercoiling both in vivo and in vitro. We have shown previously that the downstream elements of the nifL promoter are involved in the supercoiling response. In this study we find that the upstream region of nifL influences the supercoiling response of a hybrid nifL-glnAp2 promoter both in vivo and in vitro, demonstrating that the nifL upstream region also confers supercoiling sensitivity. DNA supercoiling did not appear to influence binding of NTRC to its sites in the nifL upstream region, suggesting that another function of this region, most probably DNA loop formation, is sensitive to changes in DNA topology.
Whitehall
S
S
Agricultural and Food Research Council, Institute of Plant and Science Research, University of Sussex, Brighton, UK.
Austin
S
S
Dixon
R
R
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism*; D004278:DNA, Superhelical / Q000737:chemistry / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000378:metabolism*; D009586:Nitrogen Fixation / Q000235:genetics; D009690:Nucleic Acid Conformation; D010957:Plasmids; D011196:Potassium Permanganate; D011401:Promoter Regions, Genetic / Q000235:genetics; D012808:Sigma Factor / Q000378:metabolism*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D012808:Sigma Factor; C084296:nifL protein, Bacteria; D011196:Potassium Permanganate; D012321:DNA-Directed RNA Polymerases; D001616:beta-Galactosidase