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¹ School of Biological Sciences, University of Manchester, Manchester M13 9PT, UK
² Cancer and Infection Research Area, AstraZeneca Pharmaceuticals, Mereside, Cheshire SK10 4TG, UK

Address correspondence to Stephen S. Taylor, School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Rd., Manchester M13 9PT, UK. Tel.: 44-161-275-5100. Fax: 44-161-275-5763. E-mail: stephen.taylor{at}man.ac.uk

The Aurora/Ipl1 family of protein kinases plays multiple roles in mitosis and cytokinesis. Here, we describe ZM447439, a novel selective Aurora kinase inhibitor. Cells treated with ZM447439 progress through interphase, enter mitosis normally, and assemble bipolar spindles. However, chromosome alignment, segregation, and cytokinesis all fail. Despite the presence of maloriented chromosomes, ZM447439-treated cells exit mitosis with normal kinetics, indicating that the spindle checkpoint is compromised. Indeed, ZM447439 prevents mitotic arrest after exposure to paclitaxel. RNA interference experiments suggest that these phenotypes are due to inhibition of Aurora B, not Aurora A or some other kinase. In the absence of Aurora B function, kinetochore localization of the spindle checkpoint components BubR1, Mad2, and Cenp-E is diminished. Furthermore, inhibition of Aurora B kinase activity prevents the rebinding of BubR1 to metaphase kinetochores after a reduction in centromeric tension. Aurora B kinase activity is also required for phosphorylation of BubR1 on entry into mitosis. Finally, we show that BubR1 is not only required for spindle checkpoint function, but is also required for chromosome alignment. Together, these results suggest that by targeting checkpoint proteins to kinetochores, Aurora B couples chromosome alignment with anaphase onset.

Key Words: mitosis; spindle checkpoint; chemical biology; aneuploidy; ZM447439

^* Abbreviation used in this paper: RNAi, RNA interference.

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• Slawson, C., Zachara, N. E., Vosseller, K., Cheung, W. D., Lane, M. D., Hart, G. W. (2005). Perturbations in O-linked {beta}-N-Acetylglucosamine Protein Modification Cause Severe Defects in Mitotic Progression and Cytokinesis. J. Biol. Chem. 280: 32944-32956 [Abstract] [Full Text]  
• Morrow, C. J., Tighe, A., Johnson, V. L., Scott, M. I.F., Ditchfield, C., Taylor, S. S. (2005). Bub1 and aurora B cooperate to maintain BubR1-mediated inhibition of APC/CCdc20. J. Cell Sci. 118: 3639-3652 [Abstract] [Full Text]  
• Kapoor, P., Lavoie, B. D., Frappier, L. (2005). EBP2 Plays a Key Role in Epstein-Barr Virus Mitotic Segregation and Is Regulated by Aurora Family Kinases. Mol. Cell. Biol. 25: 4934-4945 [Abstract] [Full Text]  
• Fukuda, T., Mishina, Y., Walker, M. P., DiAugustine, R. P. (2005). Conditional Transgenic System for Mouse Aurora A Kinase: Degradation by the Ubiquitin Proteasome Pathway Controls the Level of the Transgenic Protein. Mol. Cell. Biol. 25: 5270-5281 [Abstract] [Full Text]  
• Scaife, R. M. (2005). Selective and irreversible cell cycle inhibition by diphenyleneiodonium. Molecular Cancer Therapeutics 4: 876-884 [Abstract] [Full Text]  
• Regnier, V., Vagnarelli, P., Fukagawa, T., Zerjal, T., Burns, E., Trouche, D., Earnshaw, W., Brown, W. (2005). CENP-A Is Required for Accurate Chromosome Segregation and Sustained Kinetochore Association of BubR1. Mol. Cell. Biol. 25: 3967-3981 [Abstract] [Full Text]  
• Oikawa, T., Okuda, M., Ma, Z., Goorha, R., Tsujimoto, H., Inokuma, H., Fukasawa, K. (2005). Transcriptional Control of BubR1 by p53 and Suppression of Centrosome Amplification by BubR1. Mol. Cell. Biol. 25: 4046-4061 [Abstract] [Full Text]  
• Keen, N. (2005). Aurora Kinases and Cancer Therapy. aacredbook 2005: 178-181 [Full Text]  
• Mook, R. A. Jr. (2005). The Importance and Complexities of Target Class Selectivity in Drug Discovery. aacredbook 2005: 223-226 [Full Text]  
• Gadea, B. B., Ruderman, J. V. (2005). Aurora Kinase Inhibitor ZM447439 Blocks Chromosome-induced Spindle Assembly, the Completion of Chromosome Condensation, and the Establishment of the Spindle Integrity Checkpoint in Xenopus Egg Extracts. Mol. Biol. Cell 16: 1305-1318 [Abstract] [Full Text]  

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