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

BMC Molecular Biology
P JaresJ J Blow

Abstract

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. 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 ac...Continue Reading

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Citations

Aug 21, 2009·The Journal of Biological Chemistry·Irina Bruck, Daniel Kaplan
Jan 18, 2005·Nucleic Acids Research·Andrew FerenbachJ Julian Blow
Jul 17, 2008·Genes & Development·Tatsuro S TakahashiJohannes C Walter
Apr 23, 2016·Cell Cycle·James KnocklebyHoyun Lee
Apr 20, 2005·Oncogene·David Y Takeda, Anindya Dutta
Mar 2, 2006·The Journal of Biological Chemistry·Tania SilvaMartine Coue
Dec 3, 2014·The Journal of Biological Chemistry·Robert StephensonVikki M Weake
Dec 6, 2005·European Journal of Cell Biology·Baoqing GuoHoyun Lee

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Datasets Mentioned

BETA
AY460183

Methods Mentioned

BETA
PCR
immunodepletion

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