DOI: 10.1046/j.1365-2443.2000.00340.x. Central role for Cdc45 in establishing an initiation complex of DNA replication in <i>Xenopus</i> egg extracts

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

10.1046/j.1365-2443.2000.00340.x

Crossref journal-article

Wiley

Genes to Cells (311)

Abstract

BackgroundIn eukaryotes, chromosomal DNA is licensed to be replicated through the sequential loading of the origin recognition complex, Cdc6 and mini‐chromosome maintenance protein complex (MCM) onto chromatin. However, how the replication machinery is assembled onto the licensed chromatin during initiation of replication is poorly understood.ResultsUsing Xenopus egg extracts, we have investigated the role of Cdc45 in the loading of various replication proteins onto chromatin at the onset of S phase, and found that Cdc45, which required MCM for its loading, was essential for the sequential loading of replication protein A (RPA), DNA polymerase α and proliferating cell nuclear antigen (PCNA) onto chromatin. The assembly of DNA polymerase ɛ onto chromatin required Cdc45 but did not require DNA polymerase α. Analysis of nuclease‐digested chromatin fractions shows that Cdc45 formed a stable complex with either MCM or DNA polymerase α on chromatin.ConclusionsThese results demonstrate a central role for Cdc45 in activation of the licensed chromatin to form replication complexes at the onset of S phase, and suggest that Cdc45 has a dual role in the initiation of DNA replication: the unwinding of DNA and the recruiting of DNA polymerases onto DNA.

Bibliography

Mimura, S., Masuda, T., Matsui, T., & Takisawa, H. (2000). Central role for Cdc45 in establishing an initiation complex of DNA replication in Xenopus egg extracts. Genes to Cells, 5(6), 439â452. Portico.

Authors 4

Satoru Mimura (first)
Taro Masuda (additional)
Tomoko Matsui (additional)
Haruhiko Takisawa (additional)

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Dates

Type	When
Created	22 years, 5 months ago (March 11, 2003, 5:46 p.m.)
Deposited	1 year, 10 months ago (Oct. 7, 2023, 12:32 a.m.)
Indexed	1 year, 2 months ago (June 1, 2024, 1:13 a.m.)
Issued	25 years, 2 months ago (June 1, 2000)
Published	25 years, 2 months ago (June 1, 2000)
Published Online	23 years, 8 months ago (Dec. 25, 2001)
Published Print	25 years, 2 months ago (June 1, 2000)

Funders 0

None

BibTeX

@article{Mimura_2000, title={Central role for Cdc45 in establishing an initiation complex of DNA replication in Xenopus egg extracts}, volume={5}, ISSN={1365-2443}, url={http://dx.doi.org/10.1046/j.1365-2443.2000.00340.x}, DOI={10.1046/j.1365-2443.2000.00340.x}, number={6}, journal={Genes to Cells}, publisher={Wiley}, author={Mimura, Satoru and Masuda, Taro and Matsui, Tomoko and Takisawa, Haruhiko}, year={2000}, month=jun, pages={439–452} }

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