Mendez, J., Stillman, B. (November 2000) Chromatin association of human origin recognition complex, Cdc6, and minichromosome maintenance proteins during the cell cycle: Assembly of prereplication complexes in late mitosis. Molecular and Cellular Biology, 20 (22). pp. 8602-8612. ISSN 0270-7306
Abstract
Evidence obtained from studies with yeast and Xenopus indicate that the initiation of DNA replication is a multistep process. The origin recognition complex (ORC), Cdc6p, and minichromosome maintenance (MCM) proteins are required for establishing prereplication complexes, upon which initiation is triggered by the activation of cyclin-dependent kinases and the Dbf4p-dependent kinase Cdc7p. The identification of human homologues of these replication proteins allows investigation of S-phase regulation in mammalian cells. Using centrifugal elutriation of several human cell lines, we demonstrate that whereas human Orc2 (hOrc2p) and hMcm proteins are present throughout the cell cycle, hCdc6p levels vary, being very low in early G(1) and accumulating until cells enter mitosis. hCdc6p can be polyubiquitinated in vivo, and it is stabilized by proteasome inhibitors. Similar to the case for hOrc2p, a significant fraction of hCdc6p is present on chromatin throughout the cell cycle, whereas hMcm proteins alternate between soluble and chromatin-bound forms. Loading of hMcm proteins onto chromatin occurs in late mitosis concomitant with the destruction of cyclin B, indicating that the mitotic kinase activity inhibits prereplication complex formation in human cells.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | DNA-REPLICATION DNA replication S-PHASE S-Phase S Phase MCM PROTEINS MCM proteins SACCHAROMYCES-CEREVISIAE Saccharomyces Cerevisiae BUDDING YEAST Budding Yease SCHIZOSACCHAROMYCES-POMBE Pombe NUCLEAR-LOCALIZATION nuclear localization CDK PHOSPHORYLATION phosphorylation EUKARYOTIC ORIGINS eukaryotic origins DEPENDENT KINASE dependent kinase |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Chromatin dynamics bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex |
| CSHL Authors: | |
| Communities: | CSHL labs > Stillman lab |
| Highlight: | Stillman, Bruce W. |
| Depositing User: | CSHL Librarian |
| Date: | November 2000 |
| Date Deposited: | 02 Mar 2012 19:44 |
| Last Modified: | 20 Jun 2017 18:50 |
| PMCID: | PMC102165 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25021 |
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