The pol3-t hyperrecombination phenotype and DNA damage-induced recombination in Saccharomyces cerevisiae is RAD50 dependent

J Biomed Biotechnol. 2009:2009:312710. doi: 10.1155/2009/312710. Epub 2009 Oct 12.

Abstract

The DNA polymerase delta (POL3/CDC2) allele pol3-t of Saccharomyces cerevisiae has previously been shown to be sensitive to methylmethanesulfonate (MMS) and has been proposed to be involved in base excision repair. Our results, however, show that the pol3-t mutation is synergistic for MMS sensitivity with MAG1, a known base excision repair gene, but it is epistatic with rad50Delta, suggesting that POL3 may be involved not only in base excision repair but also in a RAD50 dependent function. We further studied the interaction of pol3-t with rad50Delta by examining their effect on spontaneous, MMS-, UV-, and ionizing radiation-induced intrachromosomal recombination. We found that rad50Delta completely abolishes the elevated spontaneous frequency of intrachromosomal recombination in the pol3-t mutant and significantly decreases UV- and MMS-induced recombination in both POL3 and pol3-t strains. Interestingly, rad50Delta had no effect on gamma-ray-induced recombination in both backgrounds between 0 and 50 Gy. Finally, the deletion of RAD50 had no effect on the elevated frequency of homologous integration conferred by the pol3-t mutation. RAD50 is possibly involved in resolution of replication forks that are stalled by mutagen-induced external DNA damage, or internal DNA damage produced by growing the pol3-t mutant at the restrictive temperature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Cell Survival / radiation effects
  • DNA Damage*
  • DNA Polymerase III / genetics*
  • DNA Polymerase III / metabolism
  • DNA Repair
  • DNA-Binding Proteins / genetics*
  • Dose-Response Relationship, Drug
  • Epistasis, Genetic / drug effects
  • Gamma Rays
  • Methyl Methanesulfonate / pharmacology
  • Mutagens / pharmacology
  • Mutation
  • Phenotype
  • Radiation Dosage
  • Recombination, Genetic*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics*
  • Ultraviolet Rays

Substances

  • DNA-Binding Proteins
  • Mutagens
  • RAD50 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Methyl Methanesulfonate
  • DNA Polymerase III