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Regulated communication between the upstream face of RNA polymerase and the β′ subunit jaw domain
Author(s) -
Wigneshweraraj Siva R,
Burrows Patricia C,
Nechaev Sergei,
Zenkin Nikolay,
Severinov Konstantin,
Buck Martin
Publication year - 2004
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1038/sj.emboj.7600407
Subject(s) - rna polymerase , biology , transcription (linguistics) , sigma factor , activator (genetics) , protein subunit , microbiology and biotechnology , transcription preinitiation complex , bacterial transcription , rna polymerase ii , promoter , rna , biochemistry , gene expression , gene , linguistics , philosophy
We used bacteriophage T7‐encoded transcription inhibitor gene protein 2 (gp2) as a probe to study the contribution of the Escherichia coli RNA polymerase (RNAP) β′ subunit jaw domain—the site of gp2 binding—to activator and ATP hydrolysis‐dependent open complex formation by the σ 54 ‐RNAP. We show that, unlike σ 70 ‐dependent transcription, activated transcription by σ 54 ‐RNAP is resistant to gp2. In contrast, activator and ATP hydrolysis‐independent transcription by σ 54 ‐RNAP is highly sensitive to gp2. We provide evidence that an activator‐ and ATP hydrolysis‐dependent conformational change involving the β′ jaw domain and promoter DNA is the basis for gp2‐resistant transcription by σ 54 ‐RNAP. Our results establish that accessory factors bound to the upstream face of the RNAP, communicate with the β′ jaw domain, and that such communication is subjected to regulation.