Chemical nuclease activity of 5-phenyl-1,10-phenanthroline-copper ion detects intermediates in transcription initiation by E. Coli RNA polymerase

Biochem Biophys Res Commun. 1990 Apr 30;168(2):756-62. doi: 10.1016/0006-291x(90)92386-e.

Abstract

The nuclease activity of the copper complex of 5-phenyl-1,10-phenanthroline (5-phi-OP-Cu) detects conformational changes in the lac UV-5 promoter caused by E. Coli RNA polymerase. The template strand in melted regions of initiation complexes upstream of the site of nucleotide triphosphate incorporation is very reactive. In open and elongation complexes, downstream scission sites (e.g. +4 and +5 for the open complex) on both strands are observed. The patterns of both downstream cutting sites suggest an atypical double helix at the leading edge of the transcription bubble.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA
  • DNA-Directed RNA Polymerases / genetics*
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / enzymology
  • Phenanthrolines / pharmacology*
  • Promoter Regions, Genetic / drug effects*
  • Templates, Genetic
  • Transcription, Genetic

Substances

  • Phenanthrolines
  • DNA
  • DNA-Directed RNA Polymerases
  • 1,10-phenanthroline