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Structure paper

TitleStructural basis of RNA polymerase I pre-initiation complex formation and promoter melting.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 1206, Year 2020
Publish dateMar 5, 2020
AuthorsMichael Pilsl / Christoph Engel /
PubMed AbstractTranscription of the ribosomal RNA precursor by RNA polymerase (Pol) I is a prerequisite for the biosynthesis of ribosomes in eukaryotes. Compared to Pols II and III, the mechanisms underlying ...Transcription of the ribosomal RNA precursor by RNA polymerase (Pol) I is a prerequisite for the biosynthesis of ribosomes in eukaryotes. Compared to Pols II and III, the mechanisms underlying promoter recognition, initiation complex formation and DNA melting by Pol I substantially diverge. Here, we report the high-resolution cryo-EM reconstruction of a Pol I early initiation intermediate assembled on a double-stranded promoter scaffold that prevents the establishment of downstream DNA contacts. Our analyses demonstrate how efficient promoter-backbone interaction is achieved by combined re-arrangements of flexible regions in the 'core factor' subunits Rrn7 and Rrn11. Furthermore, structure-function analysis illustrates how destabilization of the melted DNA region correlates with contraction of the polymerase cleft upon transcription activation, thereby combining promoter recruitment with DNA-melting. This suggests that molecular mechanisms and structural features of Pol I initiation have co-evolved to support the efficient melting, initial transcription and promoter clearance required for high-level rRNA synthesis.
External linksNat Commun / PubMed:32139698 / PubMed Central
MethodsEM (single particle)
Resolution3.54 Å
Structure data

10544

6tps

EMDB-10544, PDB-6tps:
early intermediate RNA Polymerase I Pre-initiation complex - eiPIC
Method: EM (single particle) / Resolution: 3.54 Å

EMDB-10663:
Focused refinement CF in eiPIC
Method: EM (single particle) / Resolution: 3.54 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • synthetic construct (others)
KeywordsTRANSCRIPTION / Transcription Initiation / Ribosome Biosynthesis / RNA Polymerase I

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