Two different mechanisms are involved for the bleomycin-induced DNA repair synthesis in permeabilized HeLa cells

Biochem Biophys Res Commun. 1989 Nov 15;164(3):1226-33. doi: 10.1016/0006-291x(89)91800-7.

Abstract

Bleomycin-induced DNA repair synthesis in the permeabilized HeLa cells was sensitive to aphidicolin, an inhibitor of DNA polymerase alpha and delta, and to dideoxythymidine triphosphate (ddTTP), a specific inhibitor of DNA polymerase beta. Upon combined treatment with these inhibitors, the DNA repair synthesis was inhibited to an even higher degree. This indicated that the aphidicolin- and ddTTP-sensitive DNA repair syntheses may occur by independent mechanisms. The structure of incomplete repair patches being accumulated in the presence of these inhibitors was investigated by digestion of DNA with exonuclease III after incubation with Klenow fragment and T4 DNA ligase. The results have suggested that the patch accumulating in the presence of aphidicolin is a single-stranded gap made by excision enzyme(s), whereas that accumulating in the presence of ddTTP may be generated by strand displacement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aphidicolin
  • Bleomycin / pharmacology*
  • Cell Membrane Permeability
  • DNA Ligases / metabolism
  • DNA Polymerase I / metabolism
  • DNA Repair / drug effects*
  • DNA Replication / drug effects*
  • Dideoxynucleotides
  • Diterpenes / pharmacology
  • HeLa Cells / drug effects
  • HeLa Cells / metabolism
  • Humans
  • Kinetics
  • Protein Biosynthesis / drug effects
  • T-Phages / enzymology
  • Thymine Nucleotides / pharmacology

Substances

  • Dideoxynucleotides
  • Diterpenes
  • Thymine Nucleotides
  • Bleomycin
  • Aphidicolin
  • DNA Polymerase I
  • DNA Ligases
  • 2',3'-dideoxythymidine triphosphate