DNA polymerase I-mediated ultraviolet repair synthesis in toluene-treated Escherichia coli

J Biol Chem. 1978 Feb 10;253(3):665-70.

Abstract

DNA synthesis after ultraviolet irradiation is low in wild type toluene-treated cells. The level of repair incorporation is greater in strains deficient in DNA polymerase I. The low level of repair synthesis is attributable to the concerted action of DNA polymerase I and polynucleotide ligase. Repair synthesis is stimulated by blocking ligase activity with the addition of nicotinamide mononucleotide (NMN) or the use of a ligase temperature-sensitive mutant. NMN stimulation is specific for DNA polymerase I-mediated repair synthesis, as it is absent in isogenic strains deficient in the polymerase function or the 5' leads to 3' exonuclease function associated with DNA polymerase I. DNA synthesis that is stimulated by NMN is proportional to the ultraviolet exposure at low doses, nonconservative in nature, and is dependent on the uvrA gene product but is independent of the recA gene product. These criteria place this synthesis in the excision repair pathway. The NMN-stimulated repair synthesis requires ATP and is N-ethylmaleimide-resistant. The use of NMN provides a direct means for evaluating the involvement of DNA polymerase I in excision repair.

Publication types

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

MeSH terms

  • DNA Polymerase I / metabolism*
  • DNA Repair*
  • DNA, Bacterial / biosynthesis
  • DNA, Bacterial / radiation effects
  • DNA-Directed DNA Polymerase / metabolism*
  • Escherichia coli / drug effects
  • Escherichia coli / metabolism*
  • Escherichia coli / radiation effects
  • Exonucleases / metabolism
  • Nicotinamide Mononucleotide / pharmacology
  • Polynucleotide Ligases / antagonists & inhibitors
  • Pyrimidine Dimers / metabolism
  • Toluene / pharmacology*
  • Ultraviolet Rays

Substances

  • DNA, Bacterial
  • Pyrimidine Dimers
  • Nicotinamide Mononucleotide
  • Toluene
  • DNA Polymerase I
  • DNA-Directed DNA Polymerase
  • Exonucleases
  • Polynucleotide Ligases