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TitleThe cryo-EM structure of a 12-subunit variant of RNA polymerase I reveals dissociation of the A49-A34.5 heterodimer and rearrangement of subunit A12.2.
Journal, issue, pagesElife, Vol. 8, Year 2019
Publish dateMar 26, 2019
AuthorsLucas Tafur / Yashar Sadian / Jonas Hanske / Rene Wetzel / Felix Weis / Christoph W Müller /
PubMed AbstractRNA polymerase (Pol) I is a 14-subunit enzyme that solely transcribes pre-ribosomal RNA. Cryo-electron microscopy (EM) structures of Pol I initiation and elongation complexes have given first ...RNA polymerase (Pol) I is a 14-subunit enzyme that solely transcribes pre-ribosomal RNA. Cryo-electron microscopy (EM) structures of Pol I initiation and elongation complexes have given first insights into the molecular mechanisms of Pol I transcription. Here, we present cryo-EM structures of yeast Pol I elongation complexes (ECs) bound to the nucleotide analog GMPCPP at 3.2 to 3.4 Å resolution that provide additional insight into the functional interplay between the Pol I-specific transcription-like factors A49-A34.5 and A12.2. Strikingly, most of the nucleotide-bound ECs lack the A49-A34.5 heterodimer and adopt a Pol II-like conformation, in which the A12.2 C-terminal domain is bound in a previously unobserved position at the A135 surface. Our structural and biochemical data suggest a mechanism where reversible binding of the A49-A34.5 heterodimer could contribute to the regulation of Pol I transcription initiation and elongation.
External linksElife / PubMed:30913026 / PubMed Central
MethodsEM (single particle)
Resolution3.18 - 4.0 Å
Structure data

0238

6hko

EMDB-0238, PDB-6hko:
Yeast RNA polymerase I elongation complex bound to nucleotide analog GMPCPP
Method: EM (single particle) / Resolution: 3.42 Å

0239

6hlq

EMDB-0239, PDB-6hlq:
Yeast RNA polymerase I* elongation complex bound to nucleotide analog GMPCPP
Method: EM (single particle) / Resolution: 3.18 Å

0240

6hlr

EMDB-0240, PDB-6hlr:
Yeast RNA polymerase I elongation complex bound to nucleotide analog GMPCPP (core focused)
Method: EM (single particle) / Resolution: 3.18 Å

0241

6hls

EMDB-0241, PDB-6hls:
Yeast apo RNA polymerase I*
Method: EM (single particle) / Resolution: 3.21 Å

EMDB-0242:
Yeast RNA polymerase I elongation complex bound to nucleotide analog GMPCPP (upstream DNA focused, unsharpened map)
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-G2P:
PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / GMP-CPP, energy-carrying molecule analogue*YM

Source
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • Saccharomyces cerevisiae (brewer's yeast)
KeywordsTRANSCRIPTION / polymerase / nucleotide / elongation / hydrolase

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