Abstract
Gene transcription is carried out by multi-subunit RNA polymerases (RNAPs). 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 cryoelectron microscopy to a unique transcription system using σ54 (σN), 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 stabilization 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 σ54.
Original language | English |
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Pages (from-to) | 1111-1120.e3 |
Journal | Molecular Cell |
Volume | 70 |
Issue number | 6 |
DOIs | |
Publication status | Published - 21 Jun 2018 |
Externally published | Yes |
Keywords
- Bacteria/genetics
- Cryoelectron Microscopy/methods
- DNA
- DNA, Bacterial/genetics
- DNA-Directed RNA Polymerases/metabolism
- Escherichia coli/genetics
- Models, Molecular
- Promoter Regions, Genetic/genetics
- Protein Binding
- Protein Conformation
- Sigma Factor/genetics
- Transcription Initiation Site/physiology
- Transcription Initiation, Genetic/physiology
- Transcription, Genetic/genetics