Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2

Ellen James, Minhao Liu, Carol Sheppard, Vladimir Mekler, Beatriz Cámara, Bing Liu, Pete Simpson, Ernesto Cota, Konstantin Severinov, Steve Matthews, Sivaramesh Wigneshweraraj

Research output: Contribution to journalArticlepeer-review

32 Citations (Scopus)

Abstract

The T7 phage-encoded small protein Gp2 is a non-DNA-binding transcription factor that interacts with the jaw domain of the Escherichia coli (Ec) RNA polymerase (RNAp) β' subunit and inhibits transcriptionally proficient promoter-complex (RPo) formation. Here, we describe the high-resolution solution structure of the Gp2-Ec β' jaw domain complex and show that Gp2 and DNA compete for binding to the β' jaw domain. We reveal that efficient inhibition of RPo formation by Gp2 requires the amino-terminal σ70 domain region 1.1 (R1.1), and that Gp2 antagonizes the obligatory movement of R1.1 during RPo formation. We demonstrate that Gp2 inhibits RPo formation not just by steric occlusion of the RNAp-DNA interaction but also through long-range antagonistic effects on RNAp-promoter interactions around the RNAp active center that likely occur due to repositioning of R1.1 by Gp2. The inhibition of Ec RNAp by Gp2 thus defines a previously uncharacterized mechanism by which bacterial transcription is regulated by a viral factor.

Original languageEnglish
Pages (from-to)755-766
Number of pages12
JournalMolecular Cell
Volume47
Issue number5
DOIs
Publication statusPublished - 14 Sep 2012
Externally publishedYes

Fingerprint

Dive into the research topics of 'Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2'. Together they form a unique fingerprint.

Cite this