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Distinct roles for cyclins E and A during DNA replication complex assembly and activation

Nature Cell Biology volume 4, pages 523–528 (2002)Cite this article

Abstract

Initiation of DNA replication is regulated by cyclin-dependent protein kinase 2 (Cdk2) in association with two different regulatory subunits, cyclin A and cyclin E (reviewed in ref. 1). But why two different cyclins are required and why their order of activation is tightly regulated are unknown. Using a cell-free system for initiation of DNA replication that is based on G1 nuclei, G1 cytosol and recombinant proteins, we find that cyclins E and A have specialized roles during the transition from G0 to S phase. Cyclin E stimulates replication complex assembly by cooperating with Cdc6, to make G1 nuclei competent to replicate in vitro. Cyclin A has two separable functions: it activates DNA synthesis by replication complexes that are already assembled, and it inhibits the assembly of new complexes. Thus, cyclin E opens a 'window of opportunity' for replication complex assembly that is closed by cyclin A. The dual functions of cyclin A ensure that the assembly phase (G1) ends before DNA synthesis (S) begins, thereby preventing re-initiation until the next cell cycle.

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Figure 1: Ability of nuclei to initiate DNA replication in S phase extracts develops as cells pass the G1 restriction point.
Figure 2: Cyclins A and E have different functions during initiation.
Figure 3: Replication complex assembly and activation in 'mid' G1 nuclei requires cyclin E before cyclin A.
Figure 4: Phosphorylation of Cdc6 blocks replication complex assembly.

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References

  1. Sherr, C. J. & Roberts, J. M. Genes Dev. 13, 1501–1512 (1999).

    Article  CAS  Google Scholar 

  2. Diffley, J. F. X. J. Cell Sci. 115, 869–872 (2001).

    Google Scholar 

  3. Blow, J. J. EMBO J. 20, 3293–3297 (2001).

    Article  CAS  Google Scholar 

  4. Seki, T. & Diffley, J. F. Proc. Natl Acad. Sci. USA 97, 14115–14120 (2000).

    Article  CAS  Google Scholar 

  5. Hanahan, D. & Weinberg, R. A. Cell 100, 57–70 (2000).

    Article  CAS  Google Scholar 

  6. Jackson, P. K., Chevalier, S., Philippe, M. & Kirschner, M. W. J. Cell Biol. 130, 755–769 (1995).

    Article  CAS  Google Scholar 

  7. Strausfield, U. P. et al. J. Cell Sci. 109, 1555–1563 (1996).

    Google Scholar 

  8. Furstenthal, L., Swanson, C., Kaiser, B. K., Eldridge, A. G. & Jackson, P. K. Nature Cell Biol. 3, 715–722 (2001).

    Article  CAS  Google Scholar 

  9. Meraldi, P., Lukas, J., Fry, A. M., Bartek, J. & Nigg, E. Nature Cell Biol. 1, 88–93 (1999).

    Article  CAS  Google Scholar 

  10. Girard, F., Strausfeld, U., Fernandez, A. & Lamb, N. J. Cell 67, 1169–1179 (1991).

    Article  CAS  Google Scholar 

  11. Pagano, M., Pepperkok, R., Verde, F., Ansorage, W. & Dreatta, G. EMBO J. 11, 961–971 (1992).

    Article  CAS  Google Scholar 

  12. Erlandsson, F., Linnman, C., Ekholm, S., Bengtsson, E. & Zetterberg, A. Exp. Cell Res. 259, 86–95 (2000).

    Article  CAS  Google Scholar 

  13. Cardoso, M. C., Leonhardt, H. & Nadal-Ginard, B. Cell 74, 1–20 (1993).

    Article  Google Scholar 

  14. Lukas, J. et al. Genes Dev. 11, 1479–1492 (1997).

    Article  CAS  Google Scholar 

  15. Resnitsky, D., Hengst, L. & Reed, S. I. Mol. cell Biol. 15, 4347–4352 (1995).

    Article  Google Scholar 

  16. Krude, T., Jackman, M., Pines, J. & Laskey, R. A. Cell 88, 109–119 (1997).

    Article  CAS  Google Scholar 

  17. Stoeber, K. et al. EMBO J. 17, 7219–7229 (1998).

    Article  CAS  Google Scholar 

  18. Petersen, B. O., Lukas, J., Sorensen, C. S., Bartek, J. & Helin, K. EMBO J. 18, 396–410 (1999).

    Article  CAS  Google Scholar 

  19. Coverley, D., Pelizon, C., Trewick, S. & Laskey, R. A. J. Cell Sci. 113, 1929–1938 (2000).

    CAS  PubMed  Google Scholar 

  20. Furstenthal, L., Kaiser, B. K., Swanson, C. & Jackson, P. K. J. Cell Biol. 152, 1267–1278 (2001).

    Article  CAS  Google Scholar 

  21. Cook, J. G. et al. Proc. Natl Acad. Sci. USA 99, 1347–1352 (2002).

    Article  CAS  Google Scholar 

  22. Pelizon, C., Madine, M. A., Romanowski, P. & Laskey, R. A. Genes Dev. 14, 2526–2533 (2000).

    Article  CAS  Google Scholar 

  23. Nguyen, V. Q., Co, C. & Li, J. J. Nature 411, 1068–1073 (2001).

    Article  CAS  Google Scholar 

  24. Sheaff, R., Groudine, M., Gordon, M., Roberts, J. & Clurman, B. Genes Dev. 11, 1464–1478 (1997).

    Article  CAS  Google Scholar 

  25. Swanson, C., Ross, J. & Jackson, P. K. Proc. Natl Acad. Sci. USA 97, 7796–7801 (2000).

    Article  CAS  Google Scholar 

  26. Dimitrova, D. S. & Gilbert, D. M. Mol. Cell 4, 963–993 (1999).

    Article  Google Scholar 

  27. Mendez, J. & Stillman, B. Mol. Cell. Biol. 20, 8602–8612 (2000).

    Article  CAS  Google Scholar 

  28. Williams, R. S., Shohet, R. V. & Stillman, B. Proc. Natl Acad. Sci. USA 94, 142–147 (1997).

    Article  CAS  Google Scholar 

  29. Madine, M. A. et al. J. Struct. Biol. 129, 198–210 (2000).

    Article  CAS  Google Scholar 

  30. Ekholm, S. V., Zickert, P., Reed, S. I. & Zetterberg, A. Mol. Cell. Biol. 21, 3256–3265 (2001).

    Article  CAS  Google Scholar 

  31. Su, T. T. & O'Farrell, P. H. J. Cell Biol. 140, 451–460 (1998).

    Article  CAS  Google Scholar 

  32. Laman, H., Coverley, D., Krude, T. K., Laskey, R. A. & Jones, N. Mol. Cell. Biol. 2, 624–635 (2001).

    Article  Google Scholar 

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Acknowledgements

We thank S. Trewick for help with Fig. 1a; T. Bradbeer for GST–cyclin-A expression vector; N. Hanlon for GST–Cdk2/GST–Cak1 co-expression vector; J. Ainscough, L. Ko-Ferrigno and M. Madine for critically reading the manuscript; and N. Jones for advice and support. This work was supported by Cancer Research UK, the Lister Institute of Preventive Medicine, the Medical Research Council and the Louis Jeantet Foundation.

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  1. Heike Laman

    Present address: Viral Oncology Laboratory, Wolfson Institute for Biomedical Research, Cruciform Building, University College, Gower Street, London, WC1E 6BT, UK

Authors and Affiliations

  1. Hutchison/MRC Research Centre, MRC Cancer Cell Unit, Hills Road, Cambridge, CB2 2XZ

    Dawn Coverley & Ronald A. Laskey

  2. Department of Biology, University of York, York, YO10 5YW, UK

    Dawn Coverley

  3. Cancer Research UK, 44 Lincoln's Inn Fields, London, WC2A 3PX

    Heike Laman

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Coverley, D., Laman, H. & Laskey, R. Distinct roles for cyclins E and A during DNA replication complex assembly and activation. Nat Cell Biol 4, 523–528 (2002). https://doi.org/10.1038/ncb813

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