A Xenopus Dbf4 homolog is required for Cdc7 chromatin binding and DNA replication

Jares, Pedro, Luciani, M. Gloria and Blow, J. Julian (2004) A Xenopus Dbf4 homolog is required for Cdc7 chromatin binding and DNA replication. BMC Molecular Biology, 5. ISSN 2661-8850

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Background Early in the cell cycle a pre-replicative complex (pre-RC) is assembled at each replication origin. This process involves the sequential assembly of the Origin Recognition Complex (ORC), Cdc6, Cdt1 and the MiniChromosome Maintenance (Mcm2-7) proteins onto chromatin to license the origin for use in the subsequent S phase. Licensed origins must then be activated by S phase-inducing cyclin-dependent kinases (S-CDKs) and the Dbf4/Cdc7 kinase. Results We have cloned a Xenopus homologue of Dbf4 (XDbf4), the sequence of which confirms the results of Furukhori et al. We have analysed the role of XDbf4 in DNA replication using cell-free extracts of Xenopus eggs. Our results indicate that XDbf4 is the regulatory subunit of XCdc7 required for DNA replication. We show that XDbf4 binds to chromatin during interphase, but unlike XCdc7, its chromatin association is independent of pre-RC formation, occurring in the absence of licensing, XCdc6 and XORC. Moreover, we show that the binding of XCdc7 to chromatin is dependent on the presence of XDbf4, whilst under certain circumstances XDbf4 can bind to chromatin in the absence of XCdc7. We provide evidence that the chromatin binding of XDbf4 that occurs in the absence of licensing depends on checkpoint activation. Conclusions We have identified XDbf4 as a functional activator of XCdc7, and show that it is required to recruit XCdc7 to chromatin. Our results also suggest that XCdc7 and XDbf4 are differentially regulated, potentially responding to different cell cycle signals.

Item Type: Article
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Depositing User: LivePure Connector
Date Deposited: 10 Jun 2024 15:31
Last Modified: 10 Jun 2024 19:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/95475
DOI: 10.1186/1471-2199-5-5

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