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

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...

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Autores principales: James, Ellen, Liu, Minhao, Sheppard, Carol, Mekler, Vladimir, Cámara, Beatriz, Liu, Bing, Simpson, Pete, Cota, Ernesto, Severinov, Konstantin, Matthews, Steve, Wigneshweraraj, Sivaramesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3778932/
https://www.ncbi.nlm.nih.gov/pubmed/22819324
http://dx.doi.org/10.1016/j.molcel.2012.06.013
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author James, Ellen
Liu, Minhao
Sheppard, Carol
Mekler, Vladimir
Cámara, Beatriz
Liu, Bing
Simpson, Pete
Cota, Ernesto
Severinov, Konstantin
Matthews, Steve
Wigneshweraraj, Sivaramesh
author_facet James, Ellen
Liu, Minhao
Sheppard, Carol
Mekler, Vladimir
Cámara, Beatriz
Liu, Bing
Simpson, Pete
Cota, Ernesto
Severinov, Konstantin
Matthews, Steve
Wigneshweraraj, Sivaramesh
author_sort James, Ellen
collection PubMed
description 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.
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spelling pubmed-37789322013-09-23 Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2 James, Ellen Liu, Minhao Sheppard, Carol Mekler, Vladimir Cámara, Beatriz Liu, Bing Simpson, Pete Cota, Ernesto Severinov, Konstantin Matthews, Steve Wigneshweraraj, Sivaramesh Mol Cell Article 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. Cell Press 2012-09-14 /pmc/articles/PMC3778932/ /pubmed/22819324 http://dx.doi.org/10.1016/j.molcel.2012.06.013 Text en © 2012 ELL & Excerpta Medica. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
spellingShingle Article
James, Ellen
Liu, Minhao
Sheppard, Carol
Mekler, Vladimir
Cámara, Beatriz
Liu, Bing
Simpson, Pete
Cota, Ernesto
Severinov, Konstantin
Matthews, Steve
Wigneshweraraj, Sivaramesh
Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2
title Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2
title_full Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2
title_fullStr Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2
title_full_unstemmed Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2
title_short Structural and Mechanistic Basis for the Inhibition of Escherichia coli RNA Polymerase by T7 Gp2
title_sort structural and mechanistic basis for the inhibition of escherichia coli rna polymerase by t7 gp2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3778932/
https://www.ncbi.nlm.nih.gov/pubmed/22819324
http://dx.doi.org/10.1016/j.molcel.2012.06.013
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