Single-molecule analysis reveals clustering and epigenetic regulation of replication origins at the yeast rDNA locus

Genes Dev. 2002 Oct 1;16(19):2479-84. doi: 10.1101/gad.232902.

Abstract

How eukaryotes specify their replication origins is an important unanswered question. Here, we analyze the replicative organization of yeast rDNA, which consists of approximately 150 identical repeats, each containing a potential origin. Using DNA combing and single-molecule imaging, we show that functional rDNA origins are clustered and interspersed with large domains where initiation is silenced. This repression is largely mediated by the Sir2p histone-deacetylase. Increased origin firing in sir2 Delta mutants leads to the accumulation of circular rDNA species, a major determinant of yeast aging. We conclude that rDNA replication is regulated epigenetically and that Sir2p may promote genome stability and longevity by suppressing replication-dependent rDNA recombination.

Publication types

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

MeSH terms

  • DNA Replication*
  • DNA, Fungal / biosynthesis*
  • DNA, Ribosomal / biosynthesis*
  • Histone Deacetylases / genetics
  • Histone Deacetylases / physiology*
  • Repetitive Sequences, Nucleic Acid*
  • Replication Origin*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / physiology
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / genetics
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / physiology*
  • Sirtuin 2
  • Sirtuins / genetics
  • Sirtuins / physiology*

Substances

  • DNA, Fungal
  • DNA, Ribosomal
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae
  • SIR2 protein, S cerevisiae
  • Sirtuin 2
  • Sirtuins
  • Histone Deacetylases