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TitleStructures of RNA Polymerase Closed and Intermediate Complexes Reveal Mechanisms of DNA Opening and Transcription Initiation.
Journal, issue, pagesMol Cell, Vol. 67, Issue 1, Page 106-116.e4, Year 2017
Publish dateJul 6, 2017
AuthorsRobert Glyde / Fuzhou Ye / Vidya Chandran Darbari / Nan Zhang / Martin Buck / Xiaodong Zhang /
PubMed AbstractGene 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 ...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 σ 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-σ 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 σ 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.
External linksMol Cell / PubMed:28579332 / PubMed Central
MethodsEM (single particle)
Resolution3.8 - 5.8 Å
Structure data

3695

5nsr

EMDB-3695, PDB-5nsr:
Cryo-EM structure of RNA polymerase-sigma54 holo enzyme with promoter DNA closed complex
Method: EM (single particle) / Resolution: 3.8 Å

3696

5nss

EMDB-3696, PDB-5nss:
Cryo-EM structure of RNA polymerase-sigma54 holoenzyme with promoter DNA and transcription activator PspF intermedate complex
Method: EM (single particle) / Resolution: 5.8 Å

EMDB-3697:
Cryo-EM structure of RNA polymerase-sigma54 holoenzyme with promoter DNA and transcription activator PspF intermedate complex
Method: EM (single particle) / Resolution: 4.9 Å

Source
  • escherichia coli k-12 (bacteria)
  • klebsiella pneumoniae (bacteria)
KeywordsTRANSCRIPTION / transcription initiation / RNA polymerase / sigma54 / transcription intermediate complex

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