Influence of Mg2+ and temperature on formation of the transcription bubble - PubMed (original) (raw)
. 1997 Jan 24;272(4):2259-67.
doi: 10.1074/jbc.272.4.2259.
Affiliations
- PMID: 8999932
- DOI: 10.1074/jbc.272.4.2259
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Influence of Mg2+ and temperature on formation of the transcription bubble
E Zaychikov et al. J Biol Chem. 1997.
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Abstract
The transcription bubble formed in the binding complex of T7A1 promoter upon Escherichia coli RNA polymerase was analyzed by chemical probes, namely by single-strand specific reagents, to map the unpaired bases in the bubble, and by FeEDTA, to analyze the accessibility of the DNA backbone. The latter probe could also be used as a local hydroxyl radical probe placed close to the Mg2+-binding site in the active center. The data show that the transcription bubble consists of two parts, an Mg2+-dependent part and an Mg2+-independent part, both having individual transition temperatures. The data further suggest that formation of a transcription active open complex is preceded by a transition state complex having enhanced affinity for those Mg2+ ions presumably participating in the formation of the catalytic site. Our data also suggests that the three catalytically active Mg2+ ions in RNA polymerase are functionally not equivalent. One/two of the three Mg2+ ions are responsible for the polymerization, the other two/one for enlargement of the transcription bubble.
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