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TitleStructural mechanism of ATP-independent transcription initiation by RNA polymerase I.
Journal, issue, pagesElife, Vol. 6, Year 2017
Publish dateJun 17, 2017
AuthorsYan Han / Chunli Yan / Thi Hoang Duong Nguyen / Ashleigh J Jackobel / Ivaylo Ivanov / Bruce A Knutson / Yuan He /
PubMed AbstractTranscription initiation by RNA Polymerase I (Pol I) depends on the Core Factor (CF) complex to recognize the upstream promoter and assemble into a Pre-Initiation Complex (PIC). Here, we solve a ...Transcription initiation by RNA Polymerase I (Pol I) depends on the Core Factor (CF) complex to recognize the upstream promoter and assemble into a Pre-Initiation Complex (PIC). Here, we solve a structure of Pol I-CF-DNA to 3.8 Å resolution using single-particle cryo-electron microscopy. The structure reveals a bipartite architecture of Core Factor and its recognition of the promoter from -27 to -16. Core Factor's intrinsic mobility correlates well with different conformational states of the Pol I cleft, in addition to the stabilization of either Rrn7 N-terminal domain near Pol I wall or the tandem winged helix domain of A49 at a partially overlapping location. Comparison of the three states in this study with the Pol II system suggests that a ratchet motion of the Core Factor-DNA sub-complex at upstream facilitates promoter melting in an ATP-independent manner, distinct from a DNA translocase actively threading the downstream DNA in the Pol II PIC.
External linksElife / PubMed:28623663 / PubMed Central
MethodsEM (single particle)
Resolution3.7 - 6.9 Å
Structure data

EMDB-8771, PDB-5w5y:
RNA polymerase I Initial Transcribing Complex
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-8772:
Core Factor local refinement from Pol I Initial Transcribing Complex at 4.2 angstrom
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-8773:
Pol I local refinement from Pol I Initial Transcribing Complex at 3.7 angstrom
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-8774, PDB-5w64:
RNA Polymerase I Initial Transcribing Complex State 1
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-8775, PDB-5w65:
RNA polymerase I Initial Transcribing Complex State 2
Method: EM (single particle) / Resolution: 4.3 Å

EMDB-8776, PDB-5w66:
RNA polymerase I Initial Transcribing Complex State 3
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-8777:
Pol I Initial Transcribing Complex with a truncated scaffold at 6.9 angstrom
Method: EM (single particle) / Resolution: 6.9 Å

Chemicals

ChemComp-ZN:
ZINC ION / Zinc

Source
  • saccharomyces cerevisiae s288c (yeast)
  • Baker's yeast (baker's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
KeywordsCryoelectron Microscopy / DNA, Fungal / Pol1 Transcription Initiation Complex Proteins / Protein Binding / RNA Polymerase I / RRN7 protein, S cerevisiae / Saccharomyces cerevisiae / Saccharomyces cerevisiae Proteins / Transcription Initiation, Genetic / TRANSCRIPTION / RNA Polymerase I Core Factor Transcription

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