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Nucleic Acids Res January 1, 2005; 33 (2): 765-75.

Licensing for DNA replication requires a strict sequential assembly of Cdc6 and Cdt1 onto chromatin in Xenopus egg extracts.

Tsuyama T , Tada S , Watanabe S , Seki M , Enomoto T .

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.

PubMed ID: 15687385
PMC ID: PMC548366
Article link: Nucleic Acids Res

Genes referenced: cdc6 cdt1 gem gmnn herpud1 mcm2 mcm4 mmut rlf

Article Images: [+] show captions
References [+] :
Bell, DNA replication in eukaryotic cells. 2002, Pubmed

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