DNA synthesis errors associated with double-strand-break repair.
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.
Strathern
J N
JN
Laboratory of Eukaryotic Gene Expression, NCI-Frederick Cancer Research and Development Center, ABL-Basic Research Program, Maryland 21702-1201, USA.
Shafer
B K
BK
McGill
C B
CB
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D015825:Chromosomes, Fungal; D003433:Crosses, Genetic; D003434:Crossing Over, Genetic; D004260:DNA Repair; D004271:DNA, Fungal / Q000096:biosynthesis*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004171:Diploidy; D016368:Frameshift Mutation; D005800:Genes, Fungal; D005838:Genotype; D006238:Haploidy; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009838:Oligodeoxyribonucleotides; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
D004271:DNA, Fungal; D009838:Oligodeoxyribonucleotides; D004259:DNA-Directed DNA Polymerase