PMID:10652363
| Citation |
Brodolin, K, Mustaev, A, Severinov, K and Nikiforov, V (2000) Identification of RNA polymerase beta' subunit segment contacting the melted region of the lacUV5 promoter. J. Biol. Chem. 275:3661-6 |
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| Abstract |
Identification of the RNA polymerase functional regions involved in interactions with promoter is a basis for understanding the mechanism of transcription initiation. We have used formaldehyde cross-linking to identify a region of Escherichia coli RNA polymerase beta' subunit contacting lacUV5 promoter in open complex. Treatment of open complex with formaldehyde results in cross-linking of beta' and sigma(70) subunits at positions -5 and -3 on the nontemplate strand of the promoter DNA. These cross-links reflect specific interactions between RNA polymerase and promoter established in open complex. The positions of formaldehyde cross-links in the beta' subunit were mapped to the N-terminal segment (Cys(198)-Met(237)), which is contiguous to the evolutionary conserved region B. The proximity of the beta' and sigma cross-links suggest that the N-terminal region of the beta' subunit, interacting with single-stranded promoter DNA, can cooperate with the sigma subunit in the process of open complex formation. |
| Links | |
| Keywords |
Amino Acid Sequence; Cross-Linking Reagents; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; DNA-Directed RNA Polymerases/genetics; DNA-Directed RNA Polymerases/metabolism; Escherichia coli; Molecular Sequence Data; Promoter Regions, Genetic/genetics |
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