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Accumulation of cyclin B1 requires E2F and cyclin-A-dependent rearrangement of the anaphase-promoting complex

Abstract

In mammalian somatic-cell cycles, progression through the G1-phase restriction point and initiation of DNA replication are controlled by the ability of the retinoblastoma tumour-suppressor protein (pRb) family to regulate the E2F/DP transcription factors1,2. Continuing transcription of E2F target genes beyond the G1/S transition is required for coordinating S-phase progression with cell division3,4,5, a process driven by cyclin-B-dependent kinase6,7 and anaphase-promoting complex (APC)-mediated proteolysis8. How E2F-dependent events at G1/S transition are orchestrated with cyclin B and APC activity remains unknown. Here, using an in vivo assay to measure protein stability in real time during the cell cycle, we show that repression of E2F activity or inhibition of cyclin-A-dependent kinase in S phase triggers the destruction of cyclin B1 through the re-assembly of APC, the ubiquitin ligase that is essential for mitotic cyclin proteolysis9, with its activatory subunit Cdh1 (refs 10,11,12,13). Phosphorylation-deficient mutant Cdh1 or immunodepletion of cyclin A resulted in assembly of active Cdh1–APC even in S-phase cells. These results implicate an E2F-dependent, cyclin A/Cdk2-mediated phosphorylation of Cdh1 in the timely accumulation of cyclin B1 and the coordination of cell-cycle progression during the post-restriction point period.

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Figure 1: Repression of E2F during S phase induces proteasome-dependent degradation of cyclin B1.
Figure 2: Cdk2 activity is essential for cyclin B1 accumulation.
Figure 3: Reactivation of pRb in S phase promotes reassociation of APC with Cdh1.
Figure 4: Phosphorylation of Cdh1 regulates Cdh1–APC assembly.

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References

  1. Dyson,N. The regulation of E2F by pRB-family proteins. Genes Dev. 12, 2245–2262 (1998).

    Article  CAS  PubMed  Google Scholar 

  2. Weinberg,R. A. The retinoblastoma protein and cell cycle control. Cell 81, 323–330 (1995).

    Article  CAS  PubMed  Google Scholar 

  3. Knudsen,E. S., Buckmaster,C., Chen,T. T., Feramisco,J. R. & Wang,J. Y. Inhibition of DNA synthesis by RB: effects on G1/S transition and S-phase progression. Genes Dev. 12, 2278–2292 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Chew,Y. P., Ellis,M., Wilkie,S. & Mittnacht,S. pRB phosphorylation mutants reveal role of pRB in regulating S phase completion by a mechanism independent of E2F. Oncogene 17, 2177–2186 (1998).

    Article  CAS  PubMed  Google Scholar 

  5. Lukas,J., Sørensen,C. S., Lukas,C., Santoni-Rugiu,E. & Bartek,J. p16INK4A, but not constitutively active pRb, can impose a sustained G1 arrest: molecular mechanism and implications for oncogenesis. Oncogene 18, 3930–3935 (1999).

    Article  CAS  PubMed  Google Scholar 

  6. Nurse,P. Universal control mechanism regulating onset of M-phase. Nature 344, 503–508 (1990).

    Article  ADS  CAS  PubMed  Google Scholar 

  7. Pines,J. & Hunter,T. Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport. J. Cell Biol. 115, 1–17 (1991).

    Article  CAS  PubMed  Google Scholar 

  8. Townsley,F. M. & Ruderman,J. V. Proteolytic ratchets that control progression through mitosis. Trends Cell Biol. 8, 238–244 (1998).

    Article  CAS  PubMed  Google Scholar 

  9. King,R. W., Deshaies,R. J., Peters,J. M. & Kirschner,M. W. How proteolysis drives the cell cycle. Science 274, 1652–1659 (1996).

    Article  ADS  CAS  PubMed  Google Scholar 

  10. Visintin,R., Prinz,S. & Amon,A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science 278, 460–463 (1997).

    Article  ADS  CAS  PubMed  Google Scholar 

  11. Zachariae,W., Schwab,M., Nasmyth,K. & Seufert,W. Control of cyclin ubiquitination by CDK-regulated binding of Hct1 to the anaphase promoting complex. Science 282, 1721–1724 (1998).

    Article  ADS  CAS  PubMed  Google Scholar 

  12. Fang,G., Yu,H. & Kirschner,M. W. Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1. Mol. Cell 2, 163–171 (1998).

    Article  CAS  PubMed  Google Scholar 

  13. Kramer,E. R., Gieffers,C., Holzl,G., Hengstschlager,M. & Peters,J. M. Activation of the human anaphase-promoting complex by proteins of the CDC20/Fizzy family. Curr. Biol. 8, 1207–1210 (1998).

    Article  CAS  PubMed  Google Scholar 

  14. Lukas,J. et al. Cyclin E-induced S phase without activation of the pRb/E2F pathway. Genes Dev. 11, 1479–1492 (1997).

    Article  CAS  PubMed  Google Scholar 

  15. Solomon,M. J., Glotzer,M., Lee,T. H., Philippe,M. & Kirschner,M. W. Cyclin activation of p34cdc2. Cell 63, 1013–1024 (1990).

    Article  CAS  PubMed  Google Scholar 

  16. Glotzer,M., Murray,A. W. & Kirschner,M. W. Cyclin is degraded by the ubiquitin pathway. Nature 349, 132–138 (1991).

    Article  ADS  CAS  PubMed  Google Scholar 

  17. Brandeis,M. & Hunt,T. The proteolysis of mitotic cyclins in mammalian cells persists from the end of mitosis until the onset of S phase. EMBO J. 15, 5280–5289 (1996).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Johnson,D. G., Cress,W. D., Jakoi,L. & Nevins,J. R. Oncogenic capacity of the E2F1 gene. Proc. Natl Acad. Sci. USA 91, 12823–12827 (1994).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  19. Wu,C.-L., Classon,M., Dyson,N. & Harlow,E. Expression of dominant-negative mutant DP-1 blocks cell cycle progression in G1. Mol. Cell Biol. 16, 3698–3706 (1994).

    Article  Google Scholar 

  20. Amon,A., Irniger,S. & Nasmyth,K. Closing the cell cycle circle in yeast: G2 cyclin proteolysis initiated at mitosis persists until the activation of G1 cyclins in the next cycle. Cell 77, 1037–1050 (1994).

    Article  CAS  PubMed  Google Scholar 

  21. Sigrist,S. J. & Lehner,C. F. Drosophila fizzy-related down-regulates mitotic cyclins and is required for cell proliferation arrest and entry into endocycles. Cell 90, 671–681 (1997).

    Article  CAS  PubMed  Google Scholar 

  22. Knoblich,J. A. et al. Cyclin E controls S phase progression and its down-regulation during Drosophila embryogenesis is required for the arrest of cell proliferation. Cell 77, 107–120 (1994).

    Article  CAS  PubMed  Google Scholar 

  23. Irniger,S. & Nasmyth,K. The anaphase-promoting complex is required in G1 arrested yeast cells to inhibit B-type cyclin accumulation and to prevent uncontrolled entry into S-phase. J. Cell Sci. 110, 1523–1531 (1997).

    Article  CAS  PubMed  Google Scholar 

  24. Nguyen,H., Gitig,D. M. & Koff,A. Cell-free degradation of p27kip1, a G1 cyclin-dependent kinase inhibitor, is dependent on CDK2 activity and the proteasome. Mol. Cell. Biol. 19, 1190–1201 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Sudakin,V. et al. The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis. Mol. Biol. Cell 6, 185–197 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. King,R. W. et al. A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B. Cell 81, 279–288 (1995).

    Article  CAS  PubMed  Google Scholar 

  27. Schwab,M., Lutum,A. S. & Seufert,W. Yeast Hct1 is a regulator of Clb2 cyclin proteolysis. Cell 90, 683–693 (1997).

    Article  CAS  PubMed  Google Scholar 

  28. Jaspersen,S., Charles,J. F. & Morgan,D. O. Inhibitory phosphorylation of the APC regulator Hct1 is controlled by the kinase Cdc28 and the phosphatase Cdc14. Curr. Biol. 9, 227–236 (1999).

    Article  CAS  PubMed  Google Scholar 

  29. Lukas,J. et al. Retinoblastoma-protein-dependent cell-cycle inhibition by the tumour suppressor p16. Nature 375, 503–506 (1995).

    Article  ADS  CAS  PubMed  Google Scholar 

  30. Lukas,J. et al. Cyclin D2 is a moderately oscillating nucleoprotein required for G1 phase progression in specific cell types. Oncogene 10, 2125–2134 (1995).

    CAS  PubMed  Google Scholar 

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Acknowledgements

We dedicated this letter to the memory of M. Strauss. We thank C. Gieffers, K. Helin, M. Pagano and C.-L. Wu for providing important reagents, M. Rohde for computer assistance, K. Hansen for help with T-lymphocyte stimulation, and the Danish Cancer Society, the Human Frontier Science Programme and the Danish Medical Research Council for grant support.

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Correspondence to Jiri Bartek.

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Lukas, C., Sørensen, C., Kramer, E. et al. Accumulation of cyclin B1 requires E2F and cyclin-A-dependent rearrangement of the anaphase-promoting complex. Nature 401, 815–818 (1999). https://doi.org/10.1038/44611

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