title: Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2.
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 σ(70) 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.
authors: MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London SW7 2AZ, UK.
authors: Ellen
authors: E
authors: James
authors: Minhao
authors: M
authors: Liu
authors: Carol
authors: C
authors: Sheppard
authors: Vladimir
authors: V
authors: Mekler
authors: Beatriz
authors: B
authors: Cámara
authors: Bing
authors: B
authors: Liu
authors: Pete
authors: P
authors: Simpson
authors: Ernesto
authors: E
authors: Cota
authors: Konstantin
authors: K
authors: Severinov
authors: Steve
authors: S
authors: Matthews
authors: Sivaramesh
authors: S
authors: Wigneshweraraj
mesh_terms: D004269:DNA, Bacterial / Q000737:chemistry / Q000187:drug effects; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D008958:Models, Molecular; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D011487:Protein Conformation; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D012097:Repressor Proteins; C120334:gene 2 product, bacteriophage T7; D012321:DNA-Directed RNA Polymerases