Melting during steady-state transcription of the rrnB P1 promoter in vivo and in vitro.
The rRNA rrnB P1 promoter was probed with the single-strand-selective reagent potassium permanganate during steady-state transcription in vitro and in vivo. In both cases, a weak but significant level of permanganate sensitivity was observed, which was not changed by treatment with rifampin. In contrast, static studies showed that rifampin strongly affects the very high level signal associated with polymerases that have used ATP and CTP as initiating nucleotides. We infer that the permanganate sensitivity associated with steady-state transcription is due to polymerases that have not yet used ATP and CTP. The slow and regulated step during rrnB P1 transcription may be the use of the initiating nucleotides to catalyze stable opening of the promoter DNA.
Ohlsen
K L
KL
University of California, Los Angeles.
Gralla
J D
JD
D004277:DNA, Single-Stranded; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D011196:Potassium Permanganate / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012335:RNA, Ribosomal / Q000235:genetics*; D012293:Rifampin / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D004277:DNA, Single-Stranded; D012335:RNA, Ribosomal; D011196:Potassium Permanganate; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin