Abstract
Cyclin A/cdk2 has a role in progression through S phase, and a large pool is also activated in G2 phase. Here we report that this G2 phase pool regulates the timing of progression into mitosis. Knock down of cyclin A by siRNA or addition of a specific cdk2 small molecule inhibitor delayed entry into mitosis by delaying cells in G2 phase. The G2 phase-delayed cells contained elevated levels of inactive cyclin B/cdk1. However, increased microtubule nucleation at the centrosomes was observed, and the centrosomes stained for markers of cyclin B/cdk1 activity. Both microtubule nucleation at the centrosomes and the phosphoprotein markers were lost with short-term treatment of the cdk1/2 inhibitor roscovitine but not the Mek1/2 inhibitor U0126. Cyclin A/cdk2 localized at the centrosomes in late G2 phase after separation of the centrosomes but before the start of prophase. Thus G2 phase cyclin A/cdk2 controls the timing of entry into mitosis by controlling the subsequent activation of cyclin B/cdk1, but also has an unexpected role in coordinating the activation of cyclin B/cdk1 at the centrosome and in the nucleus.
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References
Andrews PD, Knatko E, Moore WJ, Swedlow JR . (2003). Mitotic mechanics: the auroras come into view. Curr Opin Cell Biol 15: 672–683.
Bailly E, Pines J, Hunter T, Bornens M . (1992). Cytoplasmic accumulation of cyclin B1 in human cells: association with a detergent-resistant compartment and with the centrosome. J Cell Sci 101: 529–545.
Berthet C, Aleem E, Coppola V, Tessarollo L, Kaldis P . (2003). Cdk2 knockout mice are viable. Curr Biol 13: 1775–1785.
Blomberg I, Hoffmann I . (1999). Ectopic expression of Cdc25A accelerates the G(1)/S transition and leads to premature activation of cyclin E- and cyclin A-dependent kinases. Mol Cell Biol 19: 6183–6194.
Buendia B, Draetta G, Karsenti E . (1992). Regulation of the microtubule nucleating activity of centrosomes in Xenopus egg extracts: role of cyclin A-associated protein kinase. J Cell Biol 116: 1431–1442.
Clarke PR, Leiss D, Pagano M, Karsenti E . (1992). Cyclin A- and cyclin B-dependent protein kinases are regulated by different mechanisms in Xenopus egg extracts. EMBO J 11: 1751–1761.
De Souza CP, Ellem KA, Gabrielli BG . (2000). Centrosomal and cytoplasmic Cdc2/cyclin B1 activation precedes nuclear mitotic events. Exp Cell Res 257: 11–21.
den Elzen N, Pines J . (2001). Cyclin A is destroyed in prometaphase and can delay chromosome alignment and anaphase. J Cell Biol 153: 121–136.
Dutertre S, Cazales M, Quaranta M, Froment C, Trabut V, Dozier C et al. (2004). Phosphorylation of CDC25B by Aurora-A at the centrosome contributes to the G2-M transition. J Cell Sci 117: 2523–2531.
Fung TK, Ma HT, Poon RY . (2007). Specialized roles of the two mitotic cyclins in somatic cells: cyclin a as an activator of M phase-promoting factor. Mol Biol Cell 18: 1861–1873.
Furuno N, den Elzen N, Pines J . (1999). Human cyclin A is required for mitosis until mid prophase. J Cell Biol 147: 295–306.
Gabrielli BG, De Souza CP, Tonks ID, Clark JM, Hayward NK, Ellem KA . (1996). Cytoplasmic accumulation of cdc25B phosphatase in mitosis triggers centrosomal microtubule nucleation in HeLa cells. J Cell Sci 109: 1081–1093.
Goldstone S, Pavey S, Forrest A, Sinnamon J, Gabrielli B . (2001). Cdc25-dependent activation of cyclin A/cdk2 is blocked in G2 phase arrested cells independently of ATM/ATR. Oncogene 20: 921–932.
Gong D, Pomerening JR, Myers JW, Gustavsson C, Jones JT, Hahn AT et al. (2007). Cyclin A2 regulates nuclear-envelope breakdown and the nuclear accumulation of cyclin B1. Curr Biol 17: 85–91.
Gu Y, Rosenblatt J, Morgan DO . (1992). Cell cycle regulation of CDK2 activity by phosphorylation of Thr160 and Tyr15. EMBO J 11: 3995–4005.
Hu B, Mitra J, van den Heuvel S, Enders GH . (2001). S and G2 phase roles for Cdk2 revealed by inducible expression of a dominant-negative mutant in human cells. Mol Cell Biol 21: 2755–2766.
Jackman M, Lindon C, Nigg EA, Pines J . (2003). Active cyclin B1-Cdk1 first appears on centrosomes in prophase. Nat Cell Biol 5: 143–148.
Knoblich JA, Lehner CF . (1993). Synergistic action of Drosophila cyclins A and B during the G2-M transition. EMBO J 12: 65–74.
Kramer A, Mailand N, Lukas C, Syljuasen RG, Wilkinson CJ, Nigg EA et al. (2004). Centrosome-associated Chk1 prevents premature activation of cyclin-B-Cdk1 kinase. Nat Cell Biol 6: 884–891.
Krauer KG, Burgess A, Buck M, Flanagan J, Sculley TB, Gabrielli B . (2004). The EBNA-3 gene family proteins disrupt the G2/M checkpoint. Oncogene 23: 1342–1353.
Lindqvist A, Kallstrom H, Lundgren A, Barsoum E, Rosenthal CK . (2005). Cdc25B cooperates with Cdc25A to induce mitosis but has a unique role in activating cyclin B1-Cdk1 at the centrosome. J Cell Biol 171: 35–45.
Lukas C, Sorensen CS, Kramer E, Santoni-Rugiu E, Lindeneg C, Peters JM et al. (1999). Accumulation of cyclin B1 requires E2F and cyclin-A-dependent rearrangement of the anaphase-promoting complex. Nature 401: 815–818.
Mitra J, Enders GH . (2004). Cyclin A/Cdk2 complexes regulate activation of Cdk1 and Cdc25 phosphatases in human cells. Oncogene 23: 3361–3367.
Ortega S, Prieto I, Odajima J, Martin A, Dubus P, Sotillo R et al. (2003). Cyclin-dependent kinase 2 is essential for meiosis but not for mitotic cell division in mice. Nat Genet 35: 25–31.
Pagano M, Pepperkok R, Verde F, Ansorge W, Draetta G . (1992). Cyclin A is required at two points in the human cell cycle. EMBO J 11: 961–971.
Pines J, Hunter T . (1990). Human cyclin A is adenovirus E1A-associated protein p60 and behaves differently from cyclin B. Nature 346: 760–763.
Qiu L, Burgess A, Fairlie DP, Leonard H, Parsons PG, Gabrielli BG . (2000). Histone deacetylase inhibitors trigger a G2 checkpoint in normal cells that is defective in tumor cells. Mol Biol Cell 11: 2069–2083.
Rossomando AJ, Dent P, Sturgill TW, Marshak DR . (1994). Mitogen-activated protein kinase kinase 1 (MKK1) is negatively regulated by threonine phosphorylation. Mol Cell Biol 14: 1594–1602.
Sherr CJ . (2000). The Pezcoller lecture: cancer cell cycles revisited. Cancer Res 60: 3689–3695.
Stead E, White J, Faast R, Conn S, Goldstone S, Rathjen J et al. (2002). Pluripotent cell division cycles are driven by ectopic Cdk2, cyclin A/E and E2F activities. Oncogene 21: 8320–8333.
Tetsu O, McCormick F . (2003). Proliferation of cancer cells despite CDK2 inhibition. Cancer Cell 3: 233–245.
Acknowledgements
We thank both the National Health and Medical Research Council of Australia and Queensland Cancer Fund for funding, Dr Nicole den Elzen and Dr Rose Boutros for advice and critical reading of the paper and all other members of the Gabrielli laboratory for critical advice. HB is supported with a Lions Medical Research Fellowship and BG is supported with an NHMRC Senior Research Fellowship.
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De Boer, L., Oakes, V., Beamish, H. et al. Cyclin A/cdk2 coordinates centrosomal and nuclear mitotic events. Oncogene 27, 4261–4268 (2008). https://doi.org/10.1038/onc.2008.74
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DOI: https://doi.org/10.1038/onc.2008.74
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