Abstract
REPLICATION licensing factor (RLF) ensures that eukaryotic chromosomal DNA is replicated exactly once in each cell cycle1–4. 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 eggs4. 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.
Similar content being viewed by others
References
Blow, J. J. & Laskey, R. A. Nature 332, 546–548 (1988).
Leno, G. H., Downes, C. S. & Laskey, R. A. Cell 69, 151–158 (1992).
Coverley, D., Downes, C. S., Romanowski, P. & Laskey, R. A. J. Cell. Biol. 122, 985–992 (1993).
Blow, J. J. J. Cell Biol. 122, 993–1002 (1993).
Kubota, Y. & Takisawa, H. J. Cell Biol. 123, 1321–1331 (1993).
Vesely, J. et al. Eur. J. Biochem. 224, 771–786 (1994).
Hennessy, K. M., Clark, C. D. & Botstein, D. Genes Dev. 4, 2252–2263 (1990).
Tye, B.-K. Trends Cell Biol. 4, 160–165 (1994).
Madine, M. A., Khoo, C.-Y., Mills, A. D. & Laskey, R. A. Nature 375, 421–424 (1995).
Todorov, I. T. et al. J. Cell Sci. 107, 253–265 (1994).
Gibson, S. I., Surosky, R. T. & Tye, B. K. Molec. cell. Biol. 10, 5707–5720 (1990).
Chen, Y., Hennesy, K. M., Botstein, D. & Tye, B. Proc. natn. Acad. Sci. U.S.A. 89, 10459–10463 (1992).
Maiti, A. K. & Sinha, P. J. molec. Biol. 224, 545–558 (1992).
Thömmes, P. et al. Nucleic Acids Res. 20, 1069–1074 (1992).
Coxon, A., Maundrell, K. & Kearsey, S. E. Nucleic Acids Res. 20, 5571–5577 (1992).
Yan, H., Merchant, A. M. & Tye, B. K. Genes Dev. 7, 2149–2160 (1993).
Bucci, S. et al. Int. J. devl Biol. 37, 509–517 (1993).
Hu, B., Burkhart, R., Schulte, D., Musahl, C. & Knippers, R. Nucleic Acids Res. 21, 5289–5293 (1993).
Miyake, S. et al. Molec. Biol. Cell 4, 1003–1015 (1993).
Kimura, H., Nozaki, N. & Sugimoto, K. EMBO J. 13, 4311–4320 (1994).
Forsburg, S. L. & Nurse, P. J. Cell Sci. 107, 2779–2788 (1994).
Strausfeld, U. P. et al. Curr. Biol. 4, 876–883 (1994).
Shivji, M. K. K., Grey, S. J., Strausfeld, U. P., Wood, R. D. & Blow, J. J. Curr. Biol. 4, 1062–1068 (1994).
Blow, J. J. & Nurse, P. Cell 62, 855–862 (1990).
De Roeper, A., Smith, J. A., Watt, R. A. & Barry, J. M. Nature 265, 469–470 (1977).
Crevel, G. & Cotterill, S. EMB0 J. 10, 4361–4369 (1991).
Blow, J. J. & Laskey, R. A. Cell 47, 577–587 (1986).
Kubota, Y., Mimura, S., Nishimoto, S-I., Takisawa, H. & Nojima, H. Cell (in the press).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chong, J., Mahbubani, H., Khoo, CY. et al. Purification of an MCM-containing complex as a component of the DNA replication licensing system. Nature 375, 418–421 (1995). https://doi.org/10.1038/375418a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/375418a0
- Springer Nature Limited
This article is cited by
-
Temporal and spatial topography of cell proliferation in cancer
Nature Cell Biology (2022)
-
Expression, purification and biochemical characterization of Schizosaccharomyces pombe Mcm4, 6 and 7
BMC Biochemistry (2013)
-
RAD51- and MRE11-dependent reassembly of uncoupled CMG helicase complex at collapsed replication forks
Nature Structural & Molecular Biology (2012)
-
Minichromosome Maintenance Protein 7 is a potential therapeutic target in human cancer and a novel prognostic marker of non-small cell lung cancer
Molecular Cancer (2011)