Suppression of a new allele of the yeast RAD52 gene by overexpression of RAD51, mutations in srs2 and ccr4, or mating-type heterozygosity

Genetics. 1995 May;140(1):115-27. doi: 10.1093/genetics/140.1.115.

Abstract

The RAD52 gene of Saccharomyces cerevisiae is involved both in the recombinational repair of DNA damage and in mitotic and meiotic recombination. A new allele of rad52 has been isolated that has unusual properties. Unlike other alleles of rad52, this allele (rad52-20) is partially suppressed by an srs2 deletion; srs2 mutations normally act to suppress only rad6 and rad18 mutations. In addition, although haploid rad52-20 strains are very X-ray sensitive, diploids homozygous for this allele are only slightly X-ray sensitive and undergo normal meiosis and meiotic recombination. Because rad52-20 diploids homozygous for mating type are very X-ray sensitive, mating-type heterozygosity is acting to suppress rad52-20. Mating-type heterozygosity suppresses this allele even in haploids, because sir mutations, which result in expression of the normally silent mating-type cassettes, were identified among the extragenic revertants of rad52-20. A new allele of srs2 and alleles of the transcriptional regulatory genes ccr4 and caf1 were among the other extragenic revertants of rad52-20. Because other researchers have shown that the RAD51 and RAD52 proteins interact, RAD51 on a high copy number plasmid was tested and found to suppress the rad52-20 allele, but RAD54, 55 and 57 did not suppress. The RAD51 plasmid did not suppress rad52-1. The rad52-20 allele may encode a protein that has low affinity binding to the RAD51 protein. To test whether the selected revertants suppressed rad52-20 by elevating the expression of RAD51, an integrated RAD51-lacZ fusion was genetically crossed into each revertant. Because none of the revertants increased the level of RAD51-lacZ, the revertants must exert their effect by one or more mechanisms that are not mediated by RAD51.

Publication types

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

MeSH terms

  • Alleles*
  • DNA Helicases / genetics*
  • DNA-Binding Proteins / genetics
  • Fungal Proteins / genetics*
  • Gene Expression Regulation, Fungal* / radiation effects
  • Genes, Fungal*
  • Heterozygote
  • Histone Deacetylases*
  • Homeodomain Proteins*
  • Mating Factor
  • Peptides / genetics
  • Radiation Tolerance / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombination, Genetic*
  • Repressor Proteins / genetics
  • Reproduction
  • Ribonucleases*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae / radiation effects
  • Saccharomyces cerevisiae Proteins*
  • Sequence Deletion
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae*
  • Sirtuin 2
  • Sirtuins
  • Suppression, Genetic*
  • Trans-Activators / genetics
  • Transcription Factors / genetics*

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • Homeodomain Proteins
  • MATA2 protein, S cerevisiae
  • Peptides
  • Recombinant Fusion Proteins
  • Repressor Proteins
  • SIR1 protein, S cerevisiae
  • SIR3 protein, S cerevisiae
  • SIR4 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae
  • Trans-Activators
  • Transcription Factors
  • Mating Factor
  • CCR4 protein, S cerevisiae
  • Ribonucleases
  • POP2 protein, S cerevisiae
  • SIR2 protein, S cerevisiae
  • Sirtuin 2
  • Sirtuins
  • Histone Deacetylases
  • DNA Helicases