A beta subunit mutation disrupting the catalytic function of Escherichia coli RNA polymerase

Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6018-22. doi: 10.1073/pnas.88.14.6018.

Abstract

The substitution of the evolutionarily conserved Glu-813 for lysine in the beta subunit of RNA polymerase (RNAP) causes a partial loss of function in the assembled RNAP. In the presence of the four ribonucleoside triphosphates, the mutant RNAP displayed a decreased frequency of promoter clearance and diminished elongation rate. Both defects could be compensated by raising the ribonucleoside triphosphate concentration. In the abortive initiation reaction limited by the incomplete set of ribonucleoside triphosphates, the mutant RNAP generated aberrant patterns of products indicative of their enhanced loss from the RNAP-promoter complex. A model is proposed, attributing the multiple effect of the mutation to the malfunctioning of the RNAP active center.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Codon
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism*
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Genes, Bacterial
  • Kinetics
  • Macromolecular Substances
  • Molecular Sequence Data
  • Mutagenesis
  • Plasmids
  • Promoter Regions, Genetic
  • Restriction Mapping

Substances

  • Codon
  • Macromolecular Substances
  • DNA-Directed RNA Polymerases