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 language | English |
|---|---|
| Pages (from-to) | 3427-3436 |
| Number of pages | 10 |
| Journal | Molecular Biology of the Cell |
| Volume | 14 |
| Issue number | 8 |
| DOIs | |
| State | Published - Aug 1 2003 |
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ASJC Scopus subject areas
- Molecular Biology
- Genetics
- Cell Biology
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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 journal › Article
}
TY - JOUR
T1 - Schizosacchromyces pombe Dpb2 binds to origin DNA early in S phase and is required for chromosomal DNA replication
AU - Feng, Wenyi
AU - Rodriguez-Menocal, Luis
AU - Tolun, Gökhan
AU - D'urso, Gennaro
PY - 2003/8/1
Y1 - 2003/8/1
N2 - 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.
AB - 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.
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U2 - 10.1091/mbc.E03-02-0088
DO - 10.1091/mbc.E03-02-0088
M3 - Article
VL - 14
SP - 3427
EP - 3436
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 8
ER -