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Centrosome-associated Chk1 prevents premature activation of cyclin-B–Cdk1 kinase

Nature Cell Biology volume 6, pages 884–891 (2004)Cite this article

Abstract

Entry into mitosis occurs after activation of Cdk1, resulting in chromosome condensation in the nucleus and centrosome separation, as well as increased microtubule nucleation activity in the cytoplasm. The active cyclin-B1–Cdk1 complex first appears at the centrosome, suggesting that the centrosome may facilitate the activation of mitotic regulators required for the commitment of cells to mitosis. However, the signalling pathways involved in controlling the initial activation of Cdk1 at the centrosome remain largely unknown. Here, we show that human Chk1 kinase localizes to interphase, but not mitotic, centrosomes. Chemical inhibition of Chk1 resulted in premature centrosome separation and activation of centrosome-associated Cdk1. Forced immobilization of kinase-inactive Chk1 to centrosomes also resulted in premature Cdk1 activation. Conversely, under such conditions wild-type Chk1 impaired activation of centrosome-associated Cdk1, thereby resulting in DNA endoreplication and centrosome amplification. Activation of centrosomal Cdk1 in late prophase seemed to be mediated by cytoplasmic Cdc25B, whose activity is controlled by centrosome-associated Chk1. These results suggest that centrosome-associated Chk1 shields centrosomal Cdk1 from unscheduled activation by cytoplasmic Cdc25B, thereby contributing to proper timing of the initial steps of cell division, including mitotic spindle formation.

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Figure 1: Chk1 localizes to interphase centrosomes during unperturbed cell-cycle progression.
Figure 2: Chk1 dissociates from the centrosome at the onset of mitosis.
Figure 3: Chemical inhibition of Chk1 induces premature centrosome separation and activation of centrosome-associated Cdk1.
Figure 4: Centrosome-targeted inhibition of Chk1 enhances activation of centrosome-associated Cdk1.
Figure 5: Centrosome-tagged Chk1 specifically targets Cdc25B to the centrosome.

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Acknowledgements

We thank S. Munro for providing GFP–AKAP450ΔC-3808; K. K. Khanna and A. Khodjakov for Chk1 and γ-tubulin cDNAs, respectively; R. J. Schultz (Drug Synthesis & Chemical Branch, National Cancer Institute) for UCN-01; and Cephalon Inc. for CEP-3891, CEP-2040 and CEP-6367. We are grateful to the Danish Cancer Society, Danish Medical Research Council, European Commission and the John and Birthe Meyer Foundation, for financial support. E.A.N. and C.J.W. acknowledge support from the Max Planck Society and the Fonds der Chemischen Industrie. A.K. is Heisenberg scholar of the Deutsche Forschungsgemeinschaft.

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Authors and Affiliations

  1. Department of Cell Cycle and Cancer, Danish Cancer Society, Institute of Cancer Biology, Strandboulevarden 49, Copenhagen Ø, DK-2100, Denmark

    Alwin Krämer, Niels Mailand, Claudia Lukas, Randi G. Syljuåsen, Jiri Bartek & Jiri Lukas

  2. Medizinische Klinik und Poliklinik V, Universität Heidelberg, Hospitalstr. 3, Heidelberg, 69115, Germany

    Alwin Krämer

  3. Department of Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18a, Martinsried, D-82152, Germany

    Christopher J. Wilkinson & Erich A. Nigg

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Correspondence to Jiri Bartek.

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Krämer, A., Mailand, N., Lukas, C. et al. Centrosome-associated Chk1 prevents premature activation of cyclin-B–Cdk1 kinase. Nat Cell Biol 6, 884–891 (2004). https://doi.org/10.1038/ncb1165

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