Turning Off Transcription with Bacterial RNA Polymerase through CuAAC Click Reactions of DNA Containing 5-Ethynyluracil

Chemistry. 2018 Jun 12;24(33):8311-8314. doi: 10.1002/chem.201801757. Epub 2018 May 17.

Abstract

Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction in the major groove of DNA containing 5-ethynyluracil (UE ) with azides was used for turning off sequence-specific protein-DNA interactions. The concept was first demonstrated on switching off cleavage of short modified DNA by restriction endonuclease BamHI-HF. Finally, DNA template containing UE was used for in vitro transcription with E. coli RNA polymerase and the transcription was turned off by CuAAC with 3-azidopropane-1,2-diol or 3-azido-7-hydroxycoumarin.

Keywords: DNA; RNA polymerase; epigenetics; nucleotides; transcription.

MeSH terms

  • Alkynes / chemistry*
  • Azides / chemistry*
  • Catalysis
  • Click Chemistry
  • Copper
  • Coumarins / chemistry*
  • Cycloaddition Reaction
  • DNA / chemistry*
  • DNA-Directed RNA Polymerases / chemistry*
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / chemistry*
  • RNA, Bacterial / chemistry*
  • RNA, Bacterial / metabolism
  • Uracil / analogs & derivatives

Substances

  • 3-azido-7-hydroxycoumarin
  • Alkynes
  • Azides
  • Coumarins
  • RNA, Bacterial
  • eniluracil
  • Uracil
  • Copper
  • DNA
  • DNA-Directed RNA Polymerases