Schizosacchromyces pombe Dpb2 binds to origin DNA early in S phase and is required for chromosomal DNA replication

Wenyi Feng, Luis Rodriguez-Menocal, Gökhan Tolun, Gennaro D'urso

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Genetic evidence suggests that DNA polymerase epsilon (Pol ε) has a noncatalytic essential role during the early stages of DNA replication initiation. Herein, we report the cloning and characterization of the second largest subunit of Pol ε in fission yeast, called Dpb2. We demonstrate that Dpb2 is essential for cell viability and that a temperature-sensitive mutant of dpb2 arrests with a 1C DNA content, suggesting that Dpb2 is required for initiation of DNA replication. Using a chromatin immunoprecipitation assay, we show that Dpb2, binds preferentially to origin DNA at the beginning of S phase. We also show that the C terminus of Pol ε associates with origin DNA at the same time as Dpb2. We conclude that Dpb2 is an essential protein required for an early step in DNA replication. We propose that the primary function of Dpb2 is to facilitate assembly of the replicative complex at the start of S phase. These conclusions are based on the novel cell cycle arrest phenotype of the dpb2 mutant, on the previously uncharacterized binding of Dpb2 to replication origins, and on the observation that the essential function of Pol ε is not dependent on its DNA synthesis activity.

Original languageEnglish
Pages (from-to)3427-3436
Number of pages10
JournalMolecular Biology of the Cell
Volume14
Issue number8
DOIs
StatePublished - Aug 1 2003

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DNA Replication
S Phase
DNA
DNA Polymerase II
Replication Origin
Schizosaccharomyces
Chromatin Immunoprecipitation
Cell Cycle Checkpoints
Organism Cloning
Cell Survival
Phenotype
Temperature
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Schizosacchromyces pombe Dpb2 binds to origin DNA early in S phase and is required for chromosomal DNA replication. / Feng, Wenyi; Rodriguez-Menocal, Luis; Tolun, Gökhan; D'urso, Gennaro.

In: Molecular Biology of the Cell, Vol. 14, No. 8, 01.08.2003, p. 3427-3436.

Research output: Contribution to journalArticle

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