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• W2134927450 endingPage "2779" @default.
• W2134927450 startingPage "2767" @default.
• W2134927450 abstract "Cyclin-dependent kinases (CDKs) promote the initiation of DNA replication and prevent reinitiation before mitosis, presumably through phosphorylation of key substrates at origins of replication. In fission yeast, the p65cdc18 protein is required to initiate DNA replication and interacts with the origin recognition complex (ORC) and the p34cdc2 CDK. Here we report that p65cdc18 becomes highly phosphorylated as cells undergo the G1 --> S phase transition. This modification is dependent on p34cdc2 protein kinase activity, as well as six consensus CDK phosphorylation sites within the p65cdc18 polypeptide. Genetic interactions between cdc18+ and the S-phase cyclin cig2+ suggest that CDK-dependent phosphorylation antagonizes cdc18+ function in vivo. Using site-directed mutagenesis, we show that phosphorylation at CDK consensus sites directly targets p65cdc18 for rapid degradation and inhibits its replication activity, as strong expression of a constitutively hypophosphorylated mutant form of p65cdc18 results in large amounts of DNA over-replication in vivo. Furthermore, the over-replication phenotype produced by this mutant p65cdc18 is resistant to increased mitotic cyclin/CDK activity, a known inhibitor of over-replication. Therefore, p65cdc18 is the first example of a cellular initiation factor directly regulated in vivo by CDK-dependent phosphorylation and proteolysis. Regulation of p65cdc18 by CDK phosphorylation is likely to contribute to the CDK-driven replication switch that restricts initiation at eukaryotic origins to once per cell cycle." @default.
• W2134927450 created "2016-06-24" @default.
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• W2134927450 date "1997-11-01" @default.
• W2134927450 modified "2023-10-16" @default.
• W2134927450 title "Regulation of the replication initiator protein p65^<i>cdc18</i> by CDK phosphorylation" @default.
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• W2134927450 doi "https://doi.org/10.1101/gad.11.21.2767" @default.
• W2134927450 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/316667" @default.
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