The 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

Elife. 2019 Mar 26:8:e43204. doi: 10.7554/eLife.43204.

Abstract

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.

Keywords: RNA polymerase I; S. cerevisiae; chromosomes; elongation complex; gene expression; molecular biophysics; ribosomal RNA synthesis; structural biology; transcription regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryoelectron Microscopy*
  • Protein Conformation
  • Protein Multimerization
  • Protein Subunits / chemistry
  • Protein Subunits / metabolism
  • RNA Polymerase I / ultrastructure*
  • Saccharomyces cerevisiae / enzymology

Substances

  • Protein Subunits
  • RNA Polymerase I

Associated data

  • PDB/6HLR
  • PDB/6HKO
  • PDB/6HLQ
  • PDB/6HLS