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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Abstract
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 |
| PMCID: | PMC58709 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25133 |
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