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Structures of RNA polymerase closed and intermediate complexes reveal
mechanisms of DNA opening and transcription initiation
File(s)
Author(s)
Glyde, R
Ye, F
Darbari, V
Zhang, N
Buck, M
Type
Journal Article
Abstract
Gene transcription is carried out by RNA polymerases (RNAPs). For transcription to occur, the closed promoter complex (RPc), where DNA is double stranded, must isomerize into an open promoter complex (RPo), where the DNA is melted out into a transcription bubble and the single-stranded template DNA is delivered to the RNAP active site. Using a bacterial RNAP containing the alternative σ54 factor and cryoelectron microscopy, we determined structures of RPc and the activator-bound intermediate complex en route to RPo at 3.8 and 5.8 Å. Our structures show how RNAP-σ54 interacts with promoter DNA to initiate the DNA distortions required for transcription bubble formation, and how the activator interacts with RPc, leading to significant conformational changes in RNAP and σ54 that promote RPo formation. We propose that DNA melting is an active process initiated in RPc and that the RNAP conformations of intermediates are significantly different from that of RPc and RPo.
Date Issued
2017-07-06
Date Acceptance
2017-05-05
Citation
Molecular Cell, 2017, 67 (1), pp.106-116
URI
DOI
ISSN
1097-2765
Publisher
Elsevier (Cell Press)
Start Page
106
End Page
116
Journal / Book Title
Molecular Cell
Volume
67
Issue
1
Copyright Statement
© 2017 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Sponsor
Biotechnology and Biological Sciences Research Council (BBSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Wellcome Trust
Wellcome Trust
Medical Research Council (MRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Grant Number
BB/G001278/1
BB/H012249/1
098412/Z/12/Z
WT/099128/Z/12/Z
MR/N006828/1
BB/N007816/1
Subjects
AAA protein
DNA distortion
DNA opening
RNA polymerase
sigma factor
sigma54
transcription bubble
transcription closed complex
transcription initiation
transcription intermediate complex
Developmental Biology
06 Biological Sciences
11 Medical And Health Sciences
Publication Status
Published
Date Publish Online
2017-06-01