Conformational changes in E. coli RNA polymerase during promoter recognition

Nucleic Acids Res. 1993 Dec 11;21(24):5748-53. doi: 10.1093/nar/21.24.5748.

Abstract

We analysed complexes formed during recognition of the lacUV5 promoter by E. coli RNA polymerase using formaldehyde as a DNA-protein and protein-protein cross-linking reagent. Most of the cross-linked complexes specific for the open complex (RPO) contain the beta' subunit of RNA polymerase cross-linked with promoter DNA in the regions: -50 to -49; -5 to -10; + 5 to +8 and +18 to +21. The protein-protein cross-linking pattern of contacting subunits is the same for the RNA polymerase in solution and in RPO: there are strong sigma-beta' and beta-beta' interactions. In contrast, only beta-beta' cross-links were detected in the closed (RPC) and intermediate (RPI) complexes. In presence of lac repressor before or after formation of the RPO cross-linking pattern is similar with that of RPI (RPC) complex.

MeSH terms

  • DNA-Directed RNA Polymerases / chemistry*
  • DNA-Directed RNA Polymerases / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / enzymology*
  • Promoter Regions, Genetic*
  • Protein Conformation
  • Repressor Proteins / metabolism
  • Lösungen
  • Temperature

Substances

  • Repressor Proteins
  • Lösungen
  • DNA-Directed RNA Polymerases