Structure and mechanism of the plant RNA polymerase V

Science. 2023 Mar 24;379(6638):1209-1213. doi: 10.1126/science.adf8231. Epub 2023 Mar 9.

Abstract

In addition to the conserved RNA polymerases I to III (Pols I to III) in eukaryotes, two atypical polymerases, Pols IV and V, specifically produce noncoding RNA in the RNA-directed DNA methylation pathway in plants. Here, we report on the structures of cauliflower Pol V in the free and elongation conformations. A conserved tyrosine residue of NRPE2 stacks with a double-stranded DNA branch of the transcription bubble to potentially attenuate elongation by inducing transcription stalling. The nontemplate DNA strand is captured by NRPE2 to enhance backtracking, thereby increasing 3'-5' cleavage, which likely underpins Pol V's high fidelity. The structures also illuminate the mechanism of Pol V transcription stalling and enhanced backtracking, which may be important for Pol V's retention on chromatin to serve its function in tethering downstream factors for RNA-directed DNA methylation.

MeSH terms

  • Brassica* / enzymology
  • Catalytic Domain
  • DNA Methylation*
  • DNA, Plant / metabolism
  • DNA-Directed RNA Polymerases* / chemistry
  • DNA-Directed RNA Polymerases* / metabolism
  • Plant Proteins* / metabolism
  • Protein Conformation
  • RNA, Plant* / metabolism
  • RNA, Untranslated* / metabolism

Substances

  • DNA-Directed RNA Polymerases
  • RNA, Plant
  • Plant Proteins
  • RNA, Untranslated
  • DNA, Plant