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DNA synthesis errors associated with double-strand-break repair

J N Strathern1, B K Shafer, C B McGill

  • 1Laboratory of Eukaryotic Gene Expression, NCI-Frederick Cancer Research and Development Center, ABL-Basic Research Program, Maryland 21702-1201, USA.

Genetics
|July 1, 1995
PubMed
Summary

DNA double-strand break (DSB) repair in yeast significantly increased nearby gene mutation rates. This suggests DNA repair synthesis is error-prone compared to normal DNA replication.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Yeast Research

Background:

  • DNA double-strand breaks (DSBs) are critical DNA lesions that can arise from various sources.
  • Accurate repair of DSBs is essential for maintaining genomic integrity.
  • The error rate of DNA synthesis during DSB repair is not fully understood.

Purpose of the Study:

  • To investigate whether DNA repair synthesis following a site-specific double-strand DNA break (DSB) is more error-prone than DNA replication.
  • To quantify the impact of DSB repair on the mutation frequency of a nearby genetic locus.

Main Methods:

  • Introduction of site-specific DSBs in Saccharomyces cerevisiae using the HO endonuclease under galactose-inducible control.
  • Generation of HO-induced DSBs near revertible trp1 alleles.

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  • Quantification of trp1 allele reversion rates in induced versus uninduced cells.

    • Main Results:

    • HO-induced DSBs led to an approximate 100-fold increase in the reversion rate of nearby trp1 alleles.
    • The majority of reverted alleles were located on the chromosome that underwent DNA cleavage.
    • This indicates a high error rate during the repair synthesis process.

    Conclusions:

    • DNA repair synthesis associated with DSB repair is significantly more error-prone than genome duplication.
    • Site-specific DSBs can increase local mutation frequencies, highlighting the mutagenic potential of DSB repair pathways.
    • These findings have implications for understanding genome instability and mutagenesis.