Purification of an MCM-containing complex as a component of the DNA replication licensing system
REPLICATION licensing factor (RLF) ensures that eukaryotic chromosomal DNA is replicated exactly once in each cell cycle 1–4 . On exit from metaphase, RLF is activated and binds to or modifies chromatin. This modification (the 'licence') is required for subsequent DNA replication; the lice...
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Veröffentlicht in: | Nature (London) 1995-06, Vol.375 (6530), p.418-421 |
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Sprache: | eng |
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Zusammenfassung: | REPLICATION licensing factor (RLF) ensures that eukaryotic chromosomal DNA is replicated exactly once in each cell cycle
1–4
. 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
4
. 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 Gl and is removed during replication, consistent with its being a component of the RLF system. |
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ISSN: | 0028-0836 1476-4687 |
DOI: | 10.1038/375418a0 |