Potassium permanganate susceptibility of sigma E-RNA polymerase-promoter complexes

Gene. 1996 Oct 24;177(1-2):129-32. doi: 10.1016/0378-1119(96)00288-0.

Abstract

We used potassium permanganate (KMnO4) to identify unpaired thymidine (T) residues in promoter complexes formed by RNA polymerase (RNAP) associated with sigma E (sigma E-RNAP) from Bacillus subtilis. We found that a region of the spoIIID promoter from at least -10 to +1 becomes melted in the presence of this polymerase. In promoter complexes formed by RNAP associated with a mutant sigma E that melts promoter DNA inefficiently, we noted additional KMnO4 sensitivity at the -11 position of the spoIIID promoter. We suggest that the base pair at -11 is unpaired in both mutant and wild type (wt) complexes; however, close proximity of wt sigma E-RNAP with the T at -11 may protect it from KMnO4 attack. The absence of a close contact between the mutant sigma E-RNAP and the base at -11 may explain why this polymerase uses promoters less efficiently than wt sigma E-RNAP.

Publication types

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

MeSH terms

  • Bacillus subtilis / drug effects
  • Bacillus subtilis / genetics
  • DNA-Directed RNA Polymerases / drug effects*
  • DNA-Directed RNA Polymerases / metabolism
  • Potassium Permanganate / pharmacology*
  • Promoter Regions, Genetic / drug effects*
  • Sigma Factor / drug effects*
  • Sigma Factor / metabolism
  • Thymidine / metabolism
  • Transcription Factors / drug effects*
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • Sigma Factor
  • Transcription Factors
  • sporulation-specific sigma factors
  • Potassium Permanganate
  • DNA-Directed RNA Polymerases
  • Thymidine