Purification of an MCM-containing complex as a component of the DNA replication licensing system

Chong, James P. J., Mahbubani, Hiro M., Khoo, Chong-Yee and Blow, J. Julian (1995) Purification of an MCM-containing complex as a component of the DNA replication licensing system. Nature, 375 (6530). pp. 418-421. ISSN 0028-0836

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Abstract

Replication licensing factor (RLF) ensures that eukaryotic chromosomal DNA is replicated exactly once in each cell cycle. On exit from metaphase, RLF is activated and binds to or modifies chromatin. This modification (the 'licence') is required for subsequent DNA replication; the licence is also inactivated in the process of replication. Active RLF is not imported into the nucleus, so further DNA replication cannot occur until the DNA is relicensed by passage throught mitosis. We have developed an assay to purify RLF from Xenopus eggs. Activity resolves into two components, RLF-M and RLF-B, both of which are required for licensing. RLF-M has been purified to apparent homogeneity: it consists of three polypeptides, one of which is a Xenopus homologue of the yeast MCM3 protein. Xenopus Mcm3 associates with chomatin in G1 and is removed during replication, consistent with its being a component of the RLF system.

Item Type: Article
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Depositing User: LivePure Connector
Date Deposited: 11 Jun 2024 11:30
Last Modified: 11 Jun 2024 11:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/95581
DOI: 10.1038/375418a0

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