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Title: Structure of a bacterial RNA polymerase holoenzyme open promoter complex

Abstract

Initiation of transcription is a primary means for controlling gene expression. In bacteria, the RNA polymerase (RNAP) holoenzyme binds and unwinds promoter DNA, forming the transcription bubble of the open promoter complex (RPo). We have determined crystal structures, refined to 4.14 Å-resolution, of RPo containing Thermus aquaticus RNAP holoenzyme and promoter DNA that includes the full transcription bubble. The structures, combined with biochemical analyses, reveal key features supporting the formation and maintenance of the double-strand/single-strand DNA junction at the upstream edge of the -10 element where bubble formation initiates. The results also reveal RNAP interactions with duplex DNA just upstream of the -10 element and potential protein/DNA interactions that direct the DNA template strand into the RNAP active site. Additionally a RNA primer to yield a 4 base-pair post-translocated RNA:DNA hybrid mimics an initially transcribing complex at the point where steric clash initiates abortive initiation and σ A dissociation.

Authors:
 [1];  [1];  [2];  [3];  [1]
  1. The Rockefeller Univ., New York, NY (United States)
  2. Saint Louis Univ. School of Medicine, St. Louis, MO (United States)
  3. Univ. of Wisconsin-Madison, Madison, WI (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1223793
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
eLife
Additional Journal Information:
Journal Volume: 4; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Bae, Brian, Feklistov, Andrey, Lass-Napiorkowska, Agnieszka, Landick, Robert, and Darst, Seth A. Structure of a bacterial RNA polymerase holoenzyme open promoter complex. United States: N. p., 2015. Web. doi:10.7554/eLife.08504.
Bae, Brian, Feklistov, Andrey, Lass-Napiorkowska, Agnieszka, Landick, Robert, & Darst, Seth A. Structure of a bacterial RNA polymerase holoenzyme open promoter complex. United States. doi:10.7554/eLife.08504.
Bae, Brian, Feklistov, Andrey, Lass-Napiorkowska, Agnieszka, Landick, Robert, and Darst, Seth A. Tue . "Structure of a bacterial RNA polymerase holoenzyme open promoter complex". United States. doi:10.7554/eLife.08504. https://www.osti.gov/servlets/purl/1223793.
@article{osti_1223793,
title = {Structure of a bacterial RNA polymerase holoenzyme open promoter complex},
author = {Bae, Brian and Feklistov, Andrey and Lass-Napiorkowska, Agnieszka and Landick, Robert and Darst, Seth A.},
abstractNote = {Initiation of transcription is a primary means for controlling gene expression. In bacteria, the RNA polymerase (RNAP) holoenzyme binds and unwinds promoter DNA, forming the transcription bubble of the open promoter complex (RPo). We have determined crystal structures, refined to 4.14 Å-resolution, of RPo containing Thermus aquaticus RNAP holoenzyme and promoter DNA that includes the full transcription bubble. The structures, combined with biochemical analyses, reveal key features supporting the formation and maintenance of the double-strand/single-strand DNA junction at the upstream edge of the -10 element where bubble formation initiates. The results also reveal RNAP interactions with duplex DNA just upstream of the -10 element and potential protein/DNA interactions that direct the DNA template strand into the RNAP active site. Additionally a RNA primer to yield a 4 base-pair post-translocated RNA:DNA hybrid mimics an initially transcribing complex at the point where steric clash initiates abortive initiation and σA dissociation.},
doi = {10.7554/eLife.08504},
journal = {eLife},
issn = {2050-084X},
number = ,
volume = 4,
place = {United States},
year = {2015},
month = {9}
}

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Cited by: 33 works
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