T7 Phage Protein Gp2 Inhibits the Escherichia Coli RNA Polymerase by Antagonizing Stable DNA Strand Separation Near the Transcription Start Site

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2010 Feb 6

PMID 20133868

Citations 41

Authors

Beatriz Camara

Minhao Liu

Jonathan Reynolds

Andrey Shadrin

Bing Liu

King Kwok

Peter Simpson

Robert Weinzierl

Konstantin Severinov

Ernesto Cota

Steve Matthews

Siva R Wigneshweraraj

Affiliations

Soon will be listed here.

Abstract

Infection of Escherichia coli by the T7 phage leads to rapid and selective inhibition of the host RNA polymerase (RNAP)--a multi-subunit enzyme responsible for gene transcription--by a small ( approximately 7 kDa) phage-encoded protein called Gp2. Gp2 is also a potent inhibitor of E. coli RNAP in vitro. Here we describe the first atomic resolution structure of Gp2, which reveals a distinct run of surface-exposed negatively charged amino acid residues on one side of the molecule. Our comprehensive mutagenesis data reveal that two conserved arginine residues located on the opposite side of Gp2 are important for binding to and inhibition of RNAP. Based on a structural model of the Gp2-RNAP complex, we propose that inhibition of transcription by Gp2 involves prevention of RNAP-promoter DNA interactions required for stable DNA strand separation and maintenance of the "transcription bubble" near the transcription start site, an obligatory step in the formation of a transcriptionally competent promoter complex.

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