The roles of REV3 and RAD57 in double-strand-break-repair-induced mutagenesis of Saccharomyces cerevisiae

Genetics. 2002 Nov;162(3):1063-77.

Abstract

The DNA synthesis associated with recombinational repair of chromosomal double-strand breaks (DSBs) has a lower fidelity than normal replicative DNA synthesis. Here, we use an inverted-repeat substrate to monitor the fidelity of repair of a site-specific DSB. DSB induction made by the HO endonuclease stimulates recombination >5000-fold and is associated with a >1000-fold increase in mutagenesis of an adjacent gene. We demonstrate that most break-repair-induced mutations (BRIMs) are point mutations and have a higher proportion of frameshifts than do spontaneous mutations of the same substrate. Although the REV3 translesion DNA polymerase is not required for recombination, it introduces approximately 75% of the BRIMs and approximately 90% of the base substitution mutations. Recombinational repair of the DSB is strongly dependent upon genes of the RAD52 epistasis group; however, the residual recombinants present in rad57 mutants are associated with a 5- to 20-fold increase in BRIMs. The spectrum of mutations in rad57 mutants is similar to that seen in the wild-type strain and is similarly affected by REV3. We also find that REV3 is required for the repair of MMS-induced lesions when recombinational repair is compromised. Our data suggest that Rad55p/Rad57p help limit the generation of substrates that require pol zeta during recombination.

Publication types

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

MeSH terms

  • Adenosine Triphosphatases
  • Blotting, Southern
  • DNA Repair / physiology
  • DNA Repair Enzymes
  • DNA-Binding Proteins / physiology
  • DNA-Directed DNA Polymerase / physiology*
  • Fungal Proteins / physiology*
  • Mutation / physiology*
  • Point Mutation
  • Rad51 Recombinase
  • Rad52 DNA Repair and Recombination Protein
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / physiology*

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • RAD52 protein, S cerevisiae
  • Rad52 DNA Repair and Recombination Protein
  • Saccharomyces cerevisiae Proteins
  • RAD51 protein, S cerevisiae
  • Rad51 Recombinase
  • DNA-Directed DNA Polymerase
  • REV3 protein, S cerevisiae
  • Adenosine Triphosphatases
  • RAD57 protein, S cerevisiae
  • DNA Repair Enzymes