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TitleStructures of Bacterial RNA Polymerase Complexes Reveal the Mechanism of DNA Loading and Transcription Initiation.
Journal, issue, pagesMol Cell, Vol. 70, Issue 6, Page 1111-11120.e3, Year 2018
Publish dateJun 21, 2018
AuthorsRobert Glyde / Fuzhou Ye / Milija Jovanovic / Ioly Kotta-Loizou / Martin Buck / Xiaodong Zhang /
PubMed AbstractGene 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 ...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 σ (σ), 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 σ.
External linksMol Cell / PubMed:29932903 / PubMed Central
MethodsEM (single particle)
Resolution3.4 - 4.1 Å
Structure data

0001

6gh5

EMDB-0001, PDB-6gh5:
Cryo-EM structure of bacterial RNA polymerase-sigma54 holoenzyme transcription open complex
Method: EM (single particle) / Resolution: 3.4 Å

0002

6gh6

EMDB-0002, PDB-6gh6:
Cryo-EM structure of bacterial RNA polymerase-sigma54 holoenzyme intermediate partially loaded complex
Method: EM (single particle) / Resolution: 4.1 Å

4397

6gfw

EMDB-4397, PDB-6gfw:
Cryo-EM structure of bacterial RNA polymerase-sigma54 holoenzyme initial transcribing complex
Method: EM (single particle) / Resolution: 3.7 Å

Source
  • escherichia coli k-12 (bacteria)
  • klebsiella pneumoniae (bacteria)
  • escherichia coli (strain k12) (bacteria)
KeywordsTRANSCRIPTION / transcription initiation / DNA opening / transcription bubble / de novo RNA synthesis / open complex / DNA loading / sigma factor

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