Journal
GENE
Volume 414, Issue 1-2, Pages 32-40Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.gene.2008.02.010
Keywords
cell cycle; DNA replication; Rad53; Dun1
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The Cdc7-Dbf4 complex is a conserved serine/threonine protein kinase essential for the initiation of eukaryotic DNA replication. Although an mcm5-bob1 mutation bypasses lethality conferred by mutations in CDC7 or DBF4, the Delta cdc7 mcm5-bob1 mutant is sensitive to hydroxyurea (H-U), which induces replication stress. To elucidate the reasons for HU sensitivity conferred by deletion of CDC7, we examined the role of Cdc7-Dbf4 in the replication checkpoint. We found that in Cdc7-Dbf4-deficient cells exposed to replication stress, Rad53 remains in a hypophosphorylated form, anaphase spindle is elongated, and checkpoint-specific transcription is not induced. The hypophosphorylated Rad53 exhibits a low autophosphorylation activity, and recombinant Cdc7-Dbf4 phosphorylates Rad53 in vitro. These results suggest that Cdc7-Dbf4 is required for full activation of Rad53 in response to replication stress. (c) 2008 Elsevier B.V. All rights reserved.
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