Binding of cyclin-dependent kinases to ORC and Cdc6p regulates the chromosome replication cycle

Weinreich, M., Liang, C., Chen, H. H., Stillman, B. (September 2001) Binding of cyclin-dependent kinases to ORC and Cdc6p regulates the chromosome replication cycle. Proceedings of the National Academy of Sciences of the United States of America, 98 (20). pp. 11211-11217. ISSN 0027-8424

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DOI: 10.1073/pnas.201387198


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-CIb5p kinase from interacting with ORC, but did not prevent the Gt-specific Cdc28p-CIn2p 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.

Item Type: Paper
Uncontrolled Keywords: Amino Acid Sequence Amino Acid Substitution Anaphase Animals Binding Sites Cell Cycle physiology Cell Cycle Proteins chemistry genetics metabolism Cell Line Chromosomes Fungal genetics Cyclin-Dependent Kinases metabolism DNA Replication DNA-Binding Proteins chemistry metabolism Fungal Proteins metabolism G1 Phase Genetic Complementation Test Kinetics Mitosis Molecular Sequence Data Mutagenesis Site-Directed Origin Recognition Complex Recombinant Proteins chemistry metabolism
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
organs, tissues, organelles, cell types and functions > organelles, types and functions > mitosis
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date: September 2001
Date Deposited: 17 Feb 2012 21:05
Last Modified: 10 Sep 2019 19:10
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