Contrasting effects of Elg1-RFC and Ctf18-RFC inactivation in the absence of fully functional RFC in fission yeast

J Kim, K Robertson, KJL Mylonas, FC Gray, I Charapitsa, Stuart Andrew MacNeill

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Proliferating cell nuclear antigen loading onto DNA by replication factor C (RFC) is a key step in eukaryotic DNA replication and repair processes. In this study, the C-terminal domain (CTD) of the large subunit of fission yeast RFC is shown to be essential for its function in vivo. Cells carrying a temperature-sensitive mutation in the CTD, rfc1-44, arrest with incompletely replicated chromosomes, are sensitive to DNA damaging agents, are synthetically lethal with other DNA replication mutants, and can be suppressed by mutations in rfc5. To assess the contribution of the RFC-like complexes Elg1-RFC and Ctf18-RFC to the viability of rfc1-44, genes encoding the large subunits of these complexes have been deleted and overexpressed. Inactivation of Ctf18-RFC by the deletion of ctf18(+), dcc1(+) or ctf8(+) is lethal in an rfc1-44 background showing that full Ctf18-RFC function is required in the absence of fully functional RFC. In contrast, rfc1-44 elg1 Delta cells are viable and overproduction of Elg1 in rfc1-44 is lethal, suggesting that Elg1-RFC plays a negative role when RFC function is inhibited. Consistent with this, the deletion of elg1(+) is shown to restore viability to rfc1-44 ctf18 Delta cells.

Original languageEnglish
Pages (from-to)4078-4089
Number of pages12
JournalNucleic Acids Research
Volume33
Issue number13
DOIs
Publication statusPublished - 2005

Keywords

  • REPLICATION FACTOR-C
  • SISTER-CHROMATID COHESION
  • CELL NUCLEAR ANTIGEN
  • DNA-POLYMERASE DELTA
  • PENTAPEPTIDE SCANNING MUTAGENESIS
  • TANDEM AFFINITY PURIFICATION
  • SACCHAROMYCES-CEREVISIAE
  • SCHIZOSACCHAROMYCES-POMBE
  • GENOME STABILITY
  • S-PHASE

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