Sir2 suppresses transcription-mediated displacement of Mcm2-7 replicative helicases at the ribosomal DNA repeats

PLoS Genet. 2019 May 13;15(5):e1008138. doi: 10.1371/journal.pgen.1008138. eCollection 2019 May.

Abstract

Repetitive DNA sequences within eukaryotic heterochromatin are poorly transcribed and replicate late in S-phase. In Saccharomyces cerevisiae, the histone deacetylase Sir2 is required for both transcriptional silencing and late replication at the repetitive ribosomal DNA arrays (rDNA). Despite the widespread association between transcription and replication timing, it remains unclear how transcription might impinge on replication, or vice versa. Here we show that, when silencing of an RNA polymerase II (RNA Pol II)-transcribed non-coding RNA at the rDNA is disrupted by SIR2 deletion, RNA polymerase pushes and thereby relocalizes replicative Mcm2-7 helicases away from their loading sites to an adjacent region with low nucleosome occupancy, and this relocalization is associated with increased rDNA origin efficiency. Our results suggest a model in which two of the major defining features of heterochromatin, transcriptional silencing and late replication, are mechanistically linked through suppression of polymerase-mediated displacement of replication initiation complexes.

Publication types

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

MeSH terms

  • Cell Cycle Proteins / genetics
  • DNA Replication / genetics
  • DNA Replication / physiology
  • DNA, Ribosomal / genetics
  • DNA-Binding Proteins / genetics
  • Gene Expression Regulation, Fungal / genetics
  • Gene Silencing
  • Minichromosome Maintenance Proteins / genetics
  • Minichromosome Maintenance Proteins / metabolism*
  • RNA Polymerase I / genetics
  • RNA Polymerase II / genetics
  • RNA Polymerase II / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / genetics*
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / metabolism*
  • Sirtuin 2 / genetics*
  • Sirtuin 2 / metabolism*
  • Transcription, Genetic

Substances

  • Cell Cycle Proteins
  • DNA, Ribosomal
  • DNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae
  • RNA Polymerase II
  • RNA Polymerase I
  • SIR2 protein, S cerevisiae
  • Sirtuin 2
  • Minichromosome Maintenance Proteins