Schizosaccharomyces pombe Rtf2 mediates site-specific replication termination by inhibiting replication restart

Proc Natl Acad Sci U S A. 2009 May 12;106(19):7927-32. doi: 10.1073/pnas.0812323106. Epub 2009 Apr 23.

Abstract

Here, we identify a phylogenetically conserved Schizosaccharomyces pombe factor, named Rtf2, as a key requirement for efficient replication termination at the site-specific replication barrier RTS1. We show that Rtf2, a proliferating cell nuclear antigen-interacting protein, promotes termination at RTS1 by preventing replication restart; in the absence of Rtf2, we observe the establishment of "slow-moving" Srs2-dependent replication forks. Analysis of the pmt3 (SUMO) and rtf2 mutants establishes that pmt3 causes a reduction in RTS1 barrier activity, that rtf2 and pmt3 are nonadditive, and that pmt3 (SUMO) partly suppresses the rtf2-dependent replication restart. Our results are consistent with a model in which Rtf2 stabilizes the replication fork stalled at RTS1 until completion of DNA synthesis by a converging replication fork initiated at a flanking origin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • DNA Replication
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology*
  • Gene Expression Regulation
  • Genome, Fungal
  • Molecular Sequence Data
  • Mutation
  • Phenotype
  • Plasmids / metabolism
  • Proliferating Cell Nuclear Antigen / metabolism
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / physiology*
  • Sequence Homology, Amino Acid
  • Small Ubiquitin-Related Modifier Proteins / metabolism
  • Two-Hybrid System Techniques

Substances

  • DNA-Binding Proteins
  • Proliferating Cell Nuclear Antigen
  • Rtf2 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • Small Ubiquitin-Related Modifier Proteins

Associated data

  • GENBANK/FN376423