X-ray crystal structure of a reiterative transcription complex reveals an atypical RNA extension pathway

Proc Natl Acad Sci U S A. 2017 Aug 1;114(31):8211-8216. doi: 10.1073/pnas.1702741114. Epub 2017 Jun 26.

Abstract

Reiterative transcription is a noncanonical form of RNA synthesis in which a nucleotide specified by a single base in the DNA template is repetitively added to the nascent transcript. Here we determined the crystal structure of an RNA polymerase, the bacterial enzyme from Thermus thermophilus, engaged in reiterative transcription during transcription initiation at a promoter resembling the pyrG promoter of Bacillus subtilis The structure reveals that the reiterative transcript detours from the dedicated RNA exit channel and extends toward the main channel of the enzyme, thereby allowing RNA extension without displacement of the promoter recognition σ-factor. Nascent transcripts containing reiteratively added G residues are eventually extended by nonreiterative transcription, revealing an atypical pathway for the formation of a transcription elongation complex.

Keywords: RNA polymerase; X-ray crystal structure; reiterative transcription; transcript slippage; transcription initiation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacillus subtilis / genetics
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Crystallography, X-Ray
  • DNA-Directed RNA Polymerases / chemistry*
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Models, Molecular
  • Mutation
  • Poly G
  • Promoter Regions, Genetic
  • RNA, Bacterial / genetics*
  • RNA, Bacterial / metabolism
  • Rifampin / pharmacology
  • Thermus thermophilus / enzymology
  • Thermus thermophilus / genetics*
  • Transcription, Genetic*

Substances

  • Bacterial Proteins
  • RNA, Bacterial
  • Poly G
  • DNA-Directed RNA Polymerases
  • Rifampin

Associated data

  • PDB/5VOI
  • PDB/5VO8