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

James P. J. Chong, Hiro M. Mahbubani, Chong-Yee Khoo, J. Julian Blow

    Research output: Contribution to journalArticle

    297 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)418-421
    Number of pages4
    JournalNature
    Volume375
    Issue number6530
    DOIs
    Publication statusPublished - 1 Jun 1995

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    Licensure
    DNA Replication
    Xenopus
    Fungal Proteins
    DNA
    Metaphase
    Mitosis
    Eggs
    Chromatin
    Cell Cycle
    Peptides

    Cite this

    Chong, James P. J. ; Mahbubani, Hiro M. ; Khoo, Chong-Yee ; Blow, J. Julian. / Purification of an MCM-containing complex as a component of the DNA replication licensing system. In: Nature. 1995 ; Vol. 375, No. 6530. pp. 418-421.
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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.",
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    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; Blow, J. Julian.

    In: Nature, Vol. 375, No. 6530, 01.06.1995, p. 418-421.

    Research output: Contribution to journalArticle

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    AU - Chong, James P. J.

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