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Binding of Cyclin-Dependent Kinases to ORC and Cdc6p Regulates the Chromosome Replication Cycle
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Binding of Cyclin-Dependent Kinases to ORC and Cdc6p Regulates the Chromosome Replication Cycle
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Journal Article
Binding of Cyclin-Dependent Kinases to ORC and Cdc6p Regulates the Chromosome Replication Cycle
2001
Overview
Cdc6p and the origin recognition complex (ORC) are essential for assembly of a pre-replicative complex (preRC) at origins of replication, before the initiation of DNA synthesis. In the absence of Cdc6p, cells fail to initiate DNA replication and undergo a \"reductional\" mitosis, in which the unreplicated chromosomes are randomly segregated to the spindle poles. We show here that the cells harboring a mutation in the essential Cdc6p Walker A-box arrest in late mitosis, probably at anaphase. This cell cycle block requires either the three Cdc28p phosphorylation sites within the N terminus of Cdc6p or a short region (aa 8-17) that contains a Cy (Cyclin) interaction sequence. These same two Cdc6p mutants that allow a reductional mitosis are defective in binding Cdc28p kinase. In addition to Cdc6p, ORC also binds to cyclin-dependent kinases (CDKs). Interestingly, Sic1p, a CDK inhibitor protein, blocked the S phase-specific Cdc28p-Clb5p kinase from interacting with ORC, but did not prevent the G1-specific Cdc28p-Cln2p kinase-ORC interaction. We suggest that ORC, Cdc6p, and Sic1p bind to different CDKs in a cell cycle-dependent manner to temporally regulate events that (i) allow preRC formation after mitosis, (ii) prevent mitosis before DNA replication can occur, and (iii) promote initiation of DNA replication.
Publisher
National Academy of Sciences,National Acad Sciences,The National Academy of Sciences
Subject
/ Anaphase
/ Animals
/ Cell Cycle Proteins - chemistry
/ Cell Cycle Proteins - genetics
/ Cell Cycle Proteins - metabolism
/ Cells
/ Chromosomes, Fungal - genetics
/ Cyclin-Dependent Kinases - metabolism
/ Cyclins
/ DNA
/ DNA-Binding Proteins - chemistry
/ DNA-Binding Proteins - metabolism
/ Fungal Proteins - metabolism
/ G1 Phase
/ Genes
/ Genetic Complementation Test
/ Kinetics
/ Mitosis
/ Proteins
/ Recombinant Proteins - chemistry
/ Recombinant Proteins - metabolism
/ S Phase
/ Saccharomyces cerevisiae - cytology
/ Saccharomyces cerevisiae - genetics
/ Saccharomyces cerevisiae Proteins
/ Yeasts
