Structures of bacterial RNA polymerase complexes reveal mechanisms of DNA loading and transcription initiation
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Published version
Author(s)
Type
Journal Article
Abstract
Gene transcription is carried out by multi-subunit RNA polymerases (RNAP). Transcription initiation is a dynamic multi-step process that involves the opening of the double stranded DNA to form a transcription bubble and delivery of the template strand deep into the RNAP for RNA synthesis. Applying cryo electron microscopy to a unique transcription system using 54 (N), the major bacterial variant sigma factor, we capture a new intermediate state at 4.1 Å where promoter DNA is caught at the entrance of the RNAP cleft. Combining with new structures of the open promoter complex and an initial de novo transcribing complex at 3.4 and 3.7 Å respectively, our studies reveal the dynamics of DNA loading and mechanism of transcription bubble stabilisation that involves coordinated, large scale conformational changes of the universally conserved features within RNAP and DNA. In addition, our studies reveal a novel mechanism of strand separation by 54.
Date Issued
2018-06-21
Date Acceptance
2018-05-18
ISSN
1097-2765
Publisher
Elsevier
Start Page
1111
End Page
1120.e3
Journal / Book Title
Molecular Cell
Volume
70
Issue
6
Copyright Statement
© 2018 The authors. 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)
Medical Research Council (MRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Wellcome Trust
Biotechnology and Biological Sciences Research Council
Grant Number
BB/H012249/1
MR/N006828/1
BB/N007816/1
098412/Z/12/Z
BB/R018499/1
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Cell Biology
EM STRUCTURE DETERMINATION
ESCHERICHIA-COLI
CRYOELECTRON MICROSCOPY
CONFORMATIONAL-CHANGES
SIGMA(54) SIGMA(N)
CRYSTAL-STRUCTURES
II TRANSCRIPTION
DOWNSTREAM DNA
PROMOTER
REFINEMENT
DNA opening
RNA polymerase
bacterial enhancer-binding proteins
de novo RNA synthesis
promoter DNA
protein-DNA interactions
sigma
transcription activation
transcription bubble
transcription initiation
Bacteria
Cryoelectron Microscopy
DNA
DNA, Bacterial
DNA-Directed RNA Polymerases
Escherichia coli
Models, Molecular
Promoter Regions, Genetic
Protein Binding
Protein Conformation
Sigma Factor
Transcription Initiation Site
Transcription Initiation, Genetic
Transcription, Genetic
Bacteria
Escherichia coli
DNA-Directed RNA Polymerases
Sigma Factor
DNA
DNA, Bacterial
Cryoelectron Microscopy
Transcription, Genetic
Protein Conformation
Protein Binding
Transcription Initiation Site
Models, Molecular
Promoter Regions, Genetic
Transcription Initiation, Genetic
06 Biological Sciences
11 Medical and Health Sciences
Developmental Biology
Publication Status
Published
Date Publish Online
2018-06-21