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

doi:10.1038/375418a0

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 journal › Article › peer-review

319 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 language	English
Pages (from-to)	418-421
Number of pages	4
Journal	Nature
Volume	375
Issue number	6530
DOIs	https://doi.org/10.1038/375418a0
Publication status	Published - 1 Jun 1995

Access to Document

10.1038/375418a0

Cite this

@article{e4c0bf64d1cc4b669b57d56ecdf33372,

title = "Purification of an MCM-containing complex as a component of the DNA replication licensing system",

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.",

author = "Chong, {James P. J.} and Mahbubani, {Hiro M.} and Chong-Yee Khoo and Blow, {J. Julian}",

year = "1995",

month = jun,

day = "1",

doi = "10.1038/375418a0",

language = "English",

volume = "375",

pages = "418--421",

journal = "Nature",

issn = "0028-0836",

publisher = "Nature Publishing Group",

number = "6530",

}

TY - JOUR

T1 - Purification of an MCM-containing complex as a component of the DNA replication licensing system

AU - Chong, James P. J.

AU - Mahbubani, Hiro M.

AU - Khoo, Chong-Yee

AU - Blow, J. Julian

PY - 1995/6/1

Y1 - 1995/6/1

N2 - 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.

AB - 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.

U2 - 10.1038/375418a0

DO - 10.1038/375418a0

M3 - Article

C2 - 7760937

SN - 0028-0836

VL - 375

SP - 418

EP - 421

JO - Nature

JF - Nature

IS - 6530

ER -

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

Abstract

Access to Document

Fingerprint

Cite this