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Structure paper

TitleMolecular insight into RNA polymerase I promoter recognition and promoter melting.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 5543, Year 2019
Publish dateDec 5, 2019
AuthorsYashar Sadian / Florence Baudin / Lucas Tafur / Brice Murciano / Rene Wetzel / Felix Weis / Christoph W Müller /
PubMed AbstractRNA polymerase I (Pol I) assembles with core factor (CF) and Rrn3 on the rDNA core promoter for transcription initiation. Here, we report cryo-EM structures of closed, intermediate and open Pol I ...RNA polymerase I (Pol I) assembles with core factor (CF) and Rrn3 on the rDNA core promoter for transcription initiation. Here, we report cryo-EM structures of closed, intermediate and open Pol I initiation complexes from 2.7 to 3.7 Å resolution to visualize Pol I promoter melting and to structurally and biochemically characterize the recognition mechanism of Pol I promoter DNA. In the closed complex, double-stranded DNA runs outside the DNA-binding cleft. Rotation of CF and upstream DNA with respect to Pol I and Rrn3 results in the spontaneous loading and opening of the promoter followed by cleft closure and positioning of the Pol I A49 tandem winged helix domain (tWH) onto DNA. Conformational rearrangement of A49 tWH leads to a clash with Rrn3 to initiate complex disassembly and promoter escape. Comprehensive insight into the Pol I transcription initiation cycle allows comparisons with promoter opening by Pol II and Pol III.
External linksNat Commun / PubMed:31804486 / PubMed Central
MethodsEM (single particle)
Resolution2.7 - 4.0 Å
Structure data

10006

6rui

EMDB-10006, PDB-6rui:
RNA Polymerase I Pre-initiation complex DNA opening intermediate 2
Method: EM (single particle) / Resolution: 2.7 Å

10007

6ruo

EMDB-10007, PDB-6ruo:
RNA Polymerase I Open Complex conformation 1
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-10019:
RNA Polymerase I Open Complex conformation 2 focused refinement on CF
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-10020:
RNA Polymerase I Open Complex conformation 2 focused refinement on Pol
Method: EM (single particle) / Resolution: 2.96 Å

10038

6rwe

EMDB-10038, PDB-6rwe:
RNA Polymerase I Open Complex conformation 2
Method: EM (single particle) / Resolution: 3.0 Å

4982

6rqh

EMDB-4982, PDB-6rqh:
RNA Polymerase I Closed Conformation 1 (CC1)
Method: EM (single particle) / Resolution: 3.7 Å

4984

6rql

EMDB-4984, PDB-6rql:
RNA Polymerase I Closed Conformation 2 (CC2)
Method: EM (single particle) / Resolution: 2.9 Å

4985

6rqt

EMDB-4985, PDB-6rqt:
RNA Polymerase I-tWH-Rrn3-DNA
Method: EM (single particle) / Resolution: 4.0 Å

4987

6rrd

EMDB-4987, PDB-6rrd:
RNA Polymerase I Pre-initiation complex DNA opening intermediate 1
Method: EM (single particle) / Resolution: 3.1 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • synthetic construct (others)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
KeywordsTRANSCRIPTION / RNA Polymerase I / Pre-initiation complex / PIC / Closed Complex / CC / Core Factor / CF / Rrn3 / POL1 / tandem winged helix / tWH / Promoter escape / Open Complex / OC / DNA opening

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