DNA synthesis errors associated with double-strand-break repair

Genetics. 1995 Jul;140(3):965-72. doi: 10.1093/genetics/140.3.965.

Abstract

Repair of a site-specific double-strand DNA break (DSB) resulted in increased reversion frequency for a nearby allele. Site-specific DSBs were introduced into the genome of Saccharomyces cerevisiae by the endonuclease encoded by the HO gene. Expression of the HO gene from a galactose-inducible promoter allowed efficient DNA cleavage at a single site in large populations of cells. To determine whether the DNA synthesis associated with repair of DSBs has a higher error rate than that associated with genome duplication, HO-induced DSBs were generated 0.3 kb from revertible alleles of trp1. The reversion rate of the trp1 alleles was approximately 100-fold higher among cells that had experienced an HO cut than among uninduced cells. The reverted allele was found predominantly on the chromosome that experienced the DNA cleavage.

Publication types

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

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Base Sequence
  • Chromosomes, Fungal
  • Crosses, Genetic
  • Crossing Over, Genetic
  • DNA Repair*
  • DNA, Fungal / biosynthesis*
  • DNA-Directed DNA Polymerase / metabolism*
  • Diploidy
  • Frameshift Mutation*
  • Genes, Fungal
  • Genotype
  • Haploidy
  • Molecular Sequence Data
  • Mutagenesis
  • Oligodeoxyribonucleotides
  • Restriction Mapping
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism

Substances

  • DNA, Fungal
  • Oligodeoxyribonucleotides
  • DNA-Directed DNA Polymerase