Low levels of DNA polymerase alpha induce mitotic and meiotic instability in the ribosomal DNA gene cluster of Saccharomyces cerevisiae

PLoS Genet. 2008 Jun 27;4(6):e1000105. doi: 10.1371/journal.pgen.1000105.

Abstract

The ribosomal DNA (rDNA) genes of Saccharomyces cerevisiae are located in a tandem array of about 150 repeats. Using a diploid with markers flanking and within the rDNA array, we showed that low levels of DNA polymerase alpha elevate recombination between both homologues and sister chromatids, about five-fold in mitotic cells and 30-fold in meiotic cells. This stimulation is independent of Fob1p, a protein required for the programmed replication fork block (RFB) in the rDNA. We observed that the fob1 mutation alone significantly increased meiotic, but not mitotic, rDNA recombination, suggesting a meiosis-specific role for this protein. We found that meiotic cells with low polymerase alpha had decreased Sir2p binding and increased Spo11p-catalyzed double-strand DNA breaks in the rDNA. Furthermore, meiotic crossover interference in the rDNA is absent. These results suggest that the hyper-Rec phenotypes resulting from low levels of DNA polymerase alpha in mitosis and meiosis reflect two fundamentally different mechanisms: the increased mitotic recombination is likely due to increased double-strand DNA breaks (DSBs) resulting from Fob1p-independent stalled replication forks, whereas the hyper-Rec meiotic phenotype results from increased levels of Spo11-catalyzed DSBs in the rDNA.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromosomal Instability
  • DNA Polymerase I / genetics
  • DNA Polymerase I / metabolism*
  • DNA Topoisomerases, Type II / metabolism
  • DNA, Fungal / genetics
  • DNA, Ribosomal / genetics*
  • DNA-Binding Proteins / metabolism
  • Endodeoxyribonucleases
  • Histone Deacetylases / metabolism
  • Meiosis*
  • Mitosis*
  • Multigene Family
  • Phenotype
  • Recombination, Genetic*
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / metabolism
  • Sirtuin 2
  • Sirtuins / metabolism

Substances

  • DNA, Fungal
  • DNA, Ribosomal
  • DNA-Binding Proteins
  • FOB1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae
  • DNA Polymerase I
  • POL1 protein, S cerevisiae
  • Endodeoxyribonucleases
  • Spo11 protein, S cerevisiae
  • SIR2 protein, S cerevisiae
  • Sirtuin 2
  • Sirtuins
  • Histone Deacetylases
  • DNA Topoisomerases, Type II