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Structure paper

TitleStructure of RNA polymerase I transcribing ribosomal DNA genes.
Journal, issue, pagesNature, Vol. 540, Issue 7634, Page 607-610, Year 2016
Publish dateDec 22, 2016
AuthorsSimon Neyer / Michael Kunz / Christian Geiss / Merle Hantsche / Victor-Valentin Hodirnau / Anja Seybert / Christoph Engel / Margot P Scheffer / Patrick Cramer / Achilleas S Frangakis /
PubMed AbstractRNA polymerase I (Pol I) is a highly processive enzyme that transcribes ribosomal DNA (rDNA) and regulates growth of eukaryotic cells. Crystal structures of free Pol I from the yeast Saccharomyces ...RNA polymerase I (Pol I) is a highly processive enzyme that transcribes ribosomal DNA (rDNA) and regulates growth of eukaryotic cells. Crystal structures of free Pol I from the yeast Saccharomyces cerevisiae have revealed dimers of the enzyme stabilized by a 'connector' element and an expanded cleft containing the active centre in an inactive conformation. The central bridge helix was unfolded and a Pol-I-specific 'expander' element occupied the DNA-template-binding site. The structure of Pol I in its active transcribing conformation has yet to be determined, whereas structures of Pol II and Pol III have been solved with bound DNA template and RNA transcript. Here we report structures of active transcribing Pol I from yeast solved by two different cryo-electron microscopy approaches. A single-particle structure at 3.8 Å resolution reveals a contracted active centre cleft with bound DNA and RNA, and a narrowed pore beneath the active site that no longer holds the RNA-cleavage-stimulating domain of subunit A12.2. A structure at 29 Å resolution that was determined from cryo-electron tomograms of Pol I enzymes transcribing cellular rDNA confirms contraction of the cleft and reveals that incoming and exiting rDNA enclose an angle of around 150°. The structures suggest a model for the regulation of transcription elongation in which contracted and expanded polymerase conformations are associated with active and inactive states, respectively.
External linksNature / PubMed:27842382
MethodsEM (single particle) / EM (subtomogram averaging)
Resolution3.8 - 29.0 Å
Structure data

EMDB-4147, PDB-5m3f:
Yeast RNA polymerase I elongation complex at 3.8A
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-4148, PDB-5m3m:
Free monomeric RNA polymerase I at 4.0A resolution
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-4149:
Structure of RNA polymerase I transcribing rDNA genes
Method: EM (subtomogram averaging) / Resolution: 29.0 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-SO4:
SULFATE ION

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • Baker's yeast (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
KeywordsTRANSCRIPTION / RNA polymerase I

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