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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 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.

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  1. Blow, J. J. & Laskey, R. A. Nature 332, 546–548 (1988).

    Article  ADS  CAS  Google Scholar 

  2. Leno, G. H., Downes, C. S. & Laskey, R. A. Cell 69, 151–158 (1992).

    Article  CAS  Google Scholar 

  3. Coverley, D., Downes, C. S., Romanowski, P. & Laskey, R. A. J. Cell. Biol. 122, 985–992 (1993).

    Article  CAS  Google Scholar 

  4. Blow, J. J. J. Cell Biol. 122, 993–1002 (1993).

    Article  CAS  Google Scholar 

  5. Kubota, Y. & Takisawa, H. J. Cell Biol. 123, 1321–1331 (1993).

    Article  CAS  Google Scholar 

  6. Vesely, J. et al. Eur. J. Biochem. 224, 771–786 (1994).

    Article  CAS  Google Scholar 

  7. Hennessy, K. M., Clark, C. D. & Botstein, D. Genes Dev. 4, 2252–2263 (1990).

    Article  CAS  Google Scholar 

  8. Tye, B.-K. Trends Cell Biol. 4, 160–165 (1994).

    Article  CAS  Google Scholar 

  9. Madine, M. A., Khoo, C.-Y., Mills, A. D. & Laskey, R. A. Nature 375, 421–424 (1995).

    Article  ADS  CAS  Google Scholar 

  10. Todorov, I. T. et al. J. Cell Sci. 107, 253–265 (1994).

    CAS  PubMed  Google Scholar 

  11. Gibson, S. I., Surosky, R. T. & Tye, B. K. Molec. cell. Biol. 10, 5707–5720 (1990).

    Article  CAS  Google Scholar 

  12. Chen, Y., Hennesy, K. M., Botstein, D. & Tye, B. Proc. natn. Acad. Sci. U.S.A. 89, 10459–10463 (1992).

    Article  ADS  CAS  Google Scholar 

  13. Maiti, A. K. & Sinha, P. J. molec. Biol. 224, 545–558 (1992).

    Article  CAS  Google Scholar 

  14. Thömmes, P. et al. Nucleic Acids Res. 20, 1069–1074 (1992).

    Article  Google Scholar 

  15. Coxon, A., Maundrell, K. & Kearsey, S. E. Nucleic Acids Res. 20, 5571–5577 (1992).

    Article  CAS  Google Scholar 

  16. Yan, H., Merchant, A. M. & Tye, B. K. Genes Dev. 7, 2149–2160 (1993).

    Article  CAS  Google Scholar 

  17. Bucci, S. et al. Int. J. devl Biol. 37, 509–517 (1993).

    CAS  Google Scholar 

  18. Hu, B., Burkhart, R., Schulte, D., Musahl, C. & Knippers, R. Nucleic Acids Res. 21, 5289–5293 (1993).

    Article  CAS  Google Scholar 

  19. Miyake, S. et al. Molec. Biol. Cell 4, 1003–1015 (1993).

    Article  CAS  Google Scholar 

  20. Kimura, H., Nozaki, N. & Sugimoto, K. EMBO J. 13, 4311–4320 (1994).

    Article  CAS  Google Scholar 

  21. Forsburg, S. L. & Nurse, P. J. Cell Sci. 107, 2779–2788 (1994).

    CAS  PubMed  Google Scholar 

  22. Strausfeld, U. P. et al. Curr. Biol. 4, 876–883 (1994).

    Article  CAS  Google Scholar 

  23. Shivji, M. K. K., Grey, S. J., Strausfeld, U. P., Wood, R. D. & Blow, J. J. Curr. Biol. 4, 1062–1068 (1994).

    Article  CAS  Google Scholar 

  24. Blow, J. J. & Nurse, P. Cell 62, 855–862 (1990).

    Article  CAS  Google Scholar 

  25. De Roeper, A., Smith, J. A., Watt, R. A. & Barry, J. M. Nature 265, 469–470 (1977).

    Article  ADS  CAS  Google Scholar 

  26. Crevel, G. & Cotterill, S. EMB0 J. 10, 4361–4369 (1991).

    Article  CAS  Google Scholar 

  27. Blow, J. J. & Laskey, R. A. Cell 47, 577–587 (1986).

    Article  CAS  Google Scholar 

  28. Kubota, Y., Mimura, S., Nishimoto, S-I., Takisawa, H. & Nojima, H. Cell (in the press).

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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).

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