Licensing for DNA replication requires a strict sequential assembly of Cdc6 and Cdt1 onto chromatin in Xenopus egg extracts
| Autor: | Takashi Tsuyama, Masayuki Seki, Shusuke Tada, Takemi Enomoto, Saori Watanabe |
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| Rok vydání: | 2005 |
| Předmět: |
Cell Extracts
DNA Replication Genetics Chromosomal Proteins Non-Histone Xenopus Cell Cycle Proteins Eukaryotic DNA replication Xenopus Proteins Biology Article Chromatin Chromatin remodeling Cell biology DNA-Binding Proteins DNA replication factor CDT1 Licensing factor Control of chromosome duplication embryonic structures biology.protein Animals Origin recognition complex ChIA-PET Ovum |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 |
| DOI: | 10.1093/nar/gki226 |
| Popis: | Replication origins are licensed for a single initiation event by the loading of Mcm2-7 proteins during late mitosis and G1. Sequential associations of origin recognition complex, Cdc6 and Mcm2-7 are essential for completion of the licensing. Although Cdt1 also binds to the chromatin when the licensing reaction takes place, whether the binding is a requirement for Cdt1 to function is unclear. To analyze the relevance of the chromatin association of Cdt1, we carried out chromatin transfer experiments using either immunodepleted Xenopus egg extracts or purified proteins. Licensing assay and immunoblotting analyses indicated that Cdt1 could only license DNA replication and load Mcm2-7 onto DNA when it binds to chromatin that has already associated with Cdc6. These results provide evidence supporting that Cdc6 and Cdt1 must bind to chromatin in a strict order for DNA licensing to occur. |
| Databáze: | OpenAIRE |
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