Identification of RNA polymerase β′ subunit segment contacting the melted region of the lacUV5 promoter

被引:23
作者
Brodolin, K
Mustaev, A
Severinov, K
Nikiforov, V
机构
[1] Russian Acad Sci, Inst Mol Genet, Moscow 123182, Russia
[2] Publ Hlth Res Inst, New York, NY 10016 USA
[3] Rutgers State Univ, Waksman Inst, Piscataway, NJ 08854 USA
关键词
D O I
10.1074/jbc.275.5.3661
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 crosslinks 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.
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收藏
页码:3661 / 3666
页数:6
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