Binding of Multiple Rap1 Proteins Stimulates Chromosome Breakage Induction during DNA Replication

PLoS Genet. 2015 Aug 11;11(8):e1005283. doi: 10.1371/journal.pgen.1005283. eCollection 2015 Aug.

Abstract

Telomeres, the ends of linear eukaryotic chromosomes, have a specialized chromatin structure that provides a stable chromosomal terminus. In budding yeast Rap1 protein binds to telomeric TG repeat and negatively regulates telomere length. Here we show that binding of multiple Rap1 proteins stimulates DNA double-stranded break (DSB) induction at both telomeric and non-telomeric regions. Consistent with the role of DSB induction, Rap1 stimulates nearby recombination events in a dosage-dependent manner. Rap1 recruits Rif1 and Rif2 to telomeres, but neither Rif1 nor Rif2 is required for DSB induction. Rap1-mediated DSB induction involves replication fork progression but inactivation of checkpoint kinase Mec1 does not affect DSB induction. Rap1 tethering shortens artificially elongated telomeres in parallel with telomerase inhibition, and this telomere shortening does not require homologous recombination. These results suggest that Rap1 contributes to telomere homeostasis by promoting chromosome breakage.

Publication types

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

MeSH terms

  • Chromosomes, Fungal / genetics*
  • DNA Breaks, Double-Stranded
  • DNA Replication
  • DNA, Fungal / genetics
  • DNA, Fungal / metabolism
  • Protein Binding
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Shelterin Complex
  • Telomere Homeostasis
  • Telomere-Binding Proteins / metabolism*
  • Transcription Factors / metabolism*

Substances

  • DNA, Fungal
  • RAP1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Shelterin Complex
  • Telomere-Binding Proteins
  • Transcription Factors