Comparative analysis of the interactions of Escherichia coli sigma S and sigma 70 RNA polymerase holoenzyme with the stationary-phase-specific bolAp1 promoter.
We have investigated the interactions of Escherichia coli sigma 70 and sigma S holoenzyme RNA polymerases (E sigma S and E sigma 70) with the stationary-phase-specific bolAp1 promoter by various footprinting methods in vitro. E sigma S and E sigma 70 have been shown to transcribe the bolApl promoter in vitro. We have determined the effects of salt and holoenzyme concentrations on E sigma S and E sigma 70 open complex formation at the bolAp1 promoter in vitro. We have obtained a high-resolution hydroxyl radical (OH.) footprint of E sigma S and E sigma 70 on the bolApl promoter. The OH. footprinting data show remarkable similarities between the footprints of the heparin-resistant transcription complexes of the two holoenzymes which have the same +1 transcription start site. However, there are distinctive differences in the protection patterns in the region between -20 and -10 of the bolAp1 promoter. KMnO4 reactivity assays reveal that, at 37 degrees C, both holoenzymes produced similar but not identical patterns of reactivities.
Nguyen
L H
LH
McArdle Laboratory for Cancer Research, University of Wisconsin-Madison 53706, USA.
Burgess
R R
RR
D001483:Base Sequence; D018983:DNA Footprinting; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005798:Genes, Bacterial; D017665:Hydroxyl Radical / Q000378:metabolism; D008969:Molecular Sequence Data; D011196:Potassium Permanganate / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012808:Sigma Factor / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D012808:Sigma Factor; D011196:Potassium Permanganate; D017665:Hydroxyl Radical; D012321:DNA-Directed RNA Polymerases; D003850:Deoxyribonuclease I