Central role for Cdc45 in establishing an initiation complex of DNA replication in <i>Xenopus</i> egg extracts

Mimura, Satoru; Masuda, Taro; Matsui, Tomoko; Takisawa, Haruhiko

doi:10.1046/j.1365-2443.2000.00340.x

Cited by 124 publications

(144 citation statements)

References 42 publications

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“…Similarly, in Xenopus cell-free extracts, association of Pol ⑀ with chromatin during DNA replication can occur in the absence of Pol ␣ or replication protein A (RPA) (Mimura et al, 2000). Based on these findings, Mimura et al 2000 proposed that Pol ⑀ might associate with chromatin before synthesis of RNA primers by Pol ␣. We are currently testing whether the binding of Pol ␣ or RPA to replication origins occurs before, or is dependent on, the prior loading of Dpb2 or Pol ⑀.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, association of Pol ␣ with ARS-associated chromatin requires Dpb11, a protein that both genetically and physically interacts with Pol ⑀ (Masumoto et al, 2000). Similarly, in Xenopus cell-free extracts, association of Pol ⑀ with chromatin during DNA replication can occur in the absence of Pol ␣ or replication protein A (RPA) (Mimura et al, 2000). Based on these findings, Mimura et al 2000 proposed that Pol ⑀ might associate with chromatin before synthesis of RNA primers by Pol ␣.…”

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confidence: 99%

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Schizosacchromyces pombeDpb2 Binds to Origin DNA Early in S Phase and Is Required for Chromosomal DNA Replication

Feng

Rodriguez-Menocal

Tolun

et al. 2003

MBoC

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Genetic evidence suggests that DNA polymerase epsilon (Pol ⑀) 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 ⑀ 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 ⑀ 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 ⑀ is not dependent on its DNA synthesis activity. INTRODUCTIONChromosomal DNA replication in eukaryotic cells requires the activity of at least three DNA polymerases: ␣, ␦, and ⑀ (Waga and Stillman, 1998;Hubscher et al., 2000;Bell and Dutta, 2002). Pol ␣ is tightly associated with a primase activity capable of de novo DNA synthesis, suggesting that this enzyme is responsible for initiation of both leading and lagging strands (Waga and Stillman, 1998). Pol ␣/primase can only synthesize short primers that must then be extended by the activity of a processive DNA polymerase(s). Biochemical analysis of simian virus 40 (SV40) DNA replication, which has been extensively used as a model system for eukaryotic DNA replication, has shown that primers synthesized by Pol ␣ can be extended by Pol ␦ and that these two polymerases are sufficient for SV40 replication in vitro (Weinberg et al., 1990;Waga et al., 1994).A second processive DNA polymerase, Pol ⑀, is required for cell viability and chromosomal DNA replication in both fission (D'Urso and Nurse, 1997) and budding yeast (Morrison et al., 1990;Araki et al., 1992;Budd and Campbell, 1993). Pol ⑀ has also been implicated in both DNA repair (Jessberger et al., 1993;Wang et al., 1993;Shivji et al., 1995;Holmes, 1999) and cell cycle checkpoint control in eukaryotic cells (Navas et al., 1995). Recently, we have shown that the DNA polymerase and exonuclease domains of Pol ⑀ are dispensable for cell viability in fission yeast (Feng and D'Urso, 2001). Similar observations have been made for the evolutionarily distant yeast, Saccharomyces cerevisiae (Kesti et al., 1999;Dua et al., 2000). These findings have raised important questions regarding the essential role of this replicative enzyme in DNA synthesis. Although the N-terminal catalytic domains are dispensable, the C-terminal half of the enzyme is essential for cell viability and chromosomal replication in fission...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Schizosacchromyces pombeDpb2 Binds to Origin DNA Early in S Phase and Is Required for Chromosomal DNA Replication

Feng

Rodriguez-Menocal

Tolun

et al. 2003

MBoC

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“…3) indicate that cohesin is efficiently acetylated even under conditions in which PCNA is not recruited to chromatin, for example in the presence of the Cdk inhibitor p27. Previous work has shown that Cdk activity is required for recruitment of Cdc45, polymerase ␣, and PCNA to chromatin (36,37). This observation calls into question whether interaction of XEco2 with PCNA is important in vertebrate systems as it is in yeast.…”

Section: Xeco2mentioning

confidence: 99%

Cohesin Acetylation Promotes Sister Chromatid Cohesion Only in Association with the Replication Machinery

Song

Lafont

Chen

et al. 2012

Journal of Biological Chemistry

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Background:The conversion of cohesin to an active form requires DNA replication and acetylation of the Smc3 subunit of the complex by Eco acetyltransferases. Results: Cohesin acetylation occurs both when replication is blocked and after replication is complete but only ensures cohesion in association with the replication machinery. Conclusion: The context of cohesin acetylation is critical to cohesion establishment. Significance: Acetylation and cohesion establishment are regulated differently in yeast and vertebrates.

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“…S-CDK targets two additional proteins, Sld2 and Sld3, causing them to associate with Dpb11 and eventually resulting in the recruitment of the GINS proteins as well as DNA Pol 1 (Tanaka et al 2007;Zegerman and Diffley 2007;Muramatsu et al 2010). The subsequent activation of DNA unwinding is required for the recruitment of the lagging-strand DNA polymerases (DNA Pol a-primase and DNA Pol d (Mimura et al 2000;Heller et al 2011).…”

Section: A Model For Helicase Loadingmentioning

confidence: 99%

Helicase Loading at Chromosomal Origins of Replication

Bell

Kaguni

2013

Cold Spring Harbor Perspectives in Biology

116

109

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Loading of the replicative DNA helicase at origins of replication is of central importance in DNA replication. As the first of the replication fork proteins assemble at chromosomal origins of replication, the loaded helicase is required for the recruitment of the rest of the replication machinery. In this work, we review the current knowledge of helicase loading at Escherichia coli and eukaryotic origins of replication. In each case, this process requires both an origin recognition protein as well as one or more additional proteins. Comparison of these events shows intriguing similarities that suggest a similar underlying mechanism, as well as critical differences that likely reflect the distinct processes that regulate helicase loading in bacterial and eukaryotic cells.

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Central role for Cdc45 in establishing an initiation complex of DNA replication in Xenopus egg extracts

Cited by 124 publications

References 42 publications

Schizosacchromyces pombeDpb2 Binds to Origin DNA Early in S Phase and Is Required for Chromosomal DNA Replication

Schizosacchromyces pombeDpb2 Binds to Origin DNA Early in S Phase and Is Required for Chromosomal DNA Replication

Cohesin Acetylation Promotes Sister Chromatid Cohesion Only in Association with the Replication Machinery

Helicase Loading at Chromosomal Origins of Replication

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