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• W2113575427 startingPage "183" @default.
• W2113575427 abstract "Using cell-free extracts made from Xenopus eggs, we show that cdk2-cyclin E and A kinases play an important role in negatively regulating DNA replication. Specifically, we demonstrate that the cdk2 kinase concentration surrounding chromatin in extracts increases 200-fold once the chromatin is assembled into nuclei. Further, we find that if the cdk2–cyclin E or A concentration in egg cytosol is increased 16-fold before the addition of sperm chromatin, the chromatin fails to initiate DNA replication once assembled into nuclei. This demonstrates that cdk2–cyclin E or A can negatively regulate DNA replication. With respect to how this negative regulation occurs, we show that high levels of cdk2–cyclin E do not block the association of the protein complex ORC with sperm chromatin but do prevent association of MCM3, a protein essential for replication. Importantly, we find that MCM3 that is prebound to chromatin does not dissociate when cdk2– cyclin E levels are increased. Taken together our results strongly suggest that during the embryonic cell cycle, the low concentrations of cdk2–cyclin E present in the cytosol after mitosis and before nuclear formation allow proteins essential for potentiating DNA replication to bind to chromatin, and that the high concentration of cdk2–cyclin E within nuclei prevents MCM from reassociating with chromatin after replication. This situation could serve, in part, to limit DNA replication to a single round per cell cycle." @default.
• W2113575427 created "2016-06-24" @default.
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• W2113575427 date "1997-04-07" @default.
• W2113575427 modified "2023-10-11" @default.
• W2113575427 title "A Role for Cdk2 Kinase in Negatively Regulating DNA Replication during S Phase of the Cell Cycle" @default.
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• W2113575427 doi "https://doi.org/10.1083/jcb.137.1.183" @default.
• W2113575427 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2139856" @default.
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