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Vol. 11, No. 21,
pp. 2767-2779,
November 1, 1997
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 USA
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.
[Key Words: Fission yeast; cell cycle; S phase; cyclin-dependent kinase; over-replication]
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|
|
|
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|
|
|
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|
|
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|
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|
|
|
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|
|
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|
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|
|
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|
|
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|
|
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