PMID:8226874
| Citation |
Borukhov, S, Sagitov, V, Josaitis, CA, Gourse, RL and Goldfarb, A (1993) Two modes of transcription initiation in vitro at the rrnB P1 promoter of Escherichia coli. J. Biol. Chem. 268:23477-82 |
|---|---|
| Abstract |
The rrnB P1 promoter of Escherichia coli (starting sequence C-4-A-3-C-2-C-1-A+1-C+2-U+3-G+4) forms a binary complex with RNA polymerase that is highly unstable and requires the presence of transcription substrates ATP and CTP for stabilizing the enzyme-DNA association (Gourse, R. L. (1988) Nucleic Acids Res. 16, 9789-9809). We show that in the absence of UTP and GTP the stabilization is accomplished by short RNA oligomers synthesized in an unusual "-3-->" mode whereby the primer initiated at the +1 site presumably slips back by three nucleotides into the -3 site and is then extended yielding stable ternary complexes. By contrast, short oligomers initiated in the conventional "+1-->" mode without slippage do not exert the stabilization effect and are readily aborted from the promoter complex. The stable -3-->ternary complexes carry sigma factor but otherwise resemble elongation complexes in their high salt stability and in the fact that they are formed with a mutant RNA polymerase deficient in promoter binding. A model is proposed explaining the stability of the -3-->ternary complexes by RNA slipping into a putative "tight RNA binding site" in RNA polymerase which is normally occupied by RNA during elongation. |
| Links | |
| Keywords |
Cold Temperature; DNA-Directed RNA Polymerases/metabolism; Escherichia coli/genetics; Oligonucleotides/metabolism; Osmolar Concentration; Promoter Regions, Genetic; RNA, Ribosomal/genetics; Sigma Factor/metabolism; Templates, Genetic; Transcription, Genetic |
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