Abstract
Error-free mitosis depends on fidelity-monitoring checkpoint systems that ensure correct temporal and spatial coordination of chromosome segregation by the microtubule spindle apparatus. Defects in these checkpoint systems can lead to genomic instability, an important aspect of tumorigenesis. Here we show that the von Hippel-Lindau (VHL) tumour suppressor protein, pVHL, which is inactivated in hereditary and sporadic forms of renal cell carcinoma, localizes to the mitotic spindle in mammalian cells and its functional inactivation provokes spindle misorientation, spindle checkpoint weakening and chromosomal instability. Spindle misorientation is linked to unstable astral microtubules and is supressed by the restoration of wild-type pVHL in pVHL-deficient cells, but not in naturally-occurring VHL disease mutants that are defective in microtubule stabilization. Impaired spindle checkpoint function and chromosomal instability are the result of reduced Mad2 (mitotic arrest deficient 2) levels actuated by pVHL-inactivation and are rescued by re-expression of either Mad2 or pVHL in VHL-defective cells. An association between VHL inactivation, reduced Mad2 levels and increased aneuploidy was also found in human renal cancer, implying that the newly identified functions of pVHL in promoting proper spindle orientation and chromosomal stability probably contribute to tumour suppression.
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Acknowledgements
We thank all members of our laboratories for helpful discussions. The authors are grateful to P. Sorger and A. Musacchio for their generous gifts of reagents, R. Carazo Salas and M. Bortfeld-Miller for helping with microtubule assays, C. Azzalin and M. Patil for helping with chromosome spreads, the Light Microscopy Centre ETH Zurich, in particular G. Csucs and J. Kusch, for help with microscopy, M. Storz and S. Behnke for preparing the tissue array and the immunostaining and J. Schelldorfer for advise in statistical data analysis. C. R. T. and A. T. are members of the Life Science Zurich Graduate School Zurich, Program in Molecular Life Sciences. P. M. is recipient of a SNF Assistant Professorship and is supported by the Swiss National Science Foundation. W. K. is supported by a grant from the Swiss National Science Foundation.
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C.R.T., A.T., P.M. and W.K. concieved and planned the project and performed data analyses; K.L.G. contributed to microtubule polymerization measurements; S.P.R. contributed to analysis of mitotic slippage in renal cancer cells; I.J.F. generated biochemical data in VHL-deficient MEFs; P.S. and H.M. provided data on human renal cancer; A.H. observed mitotic localization of pVHL and the manuscript was prepared by C.R.T. and W.K. and edited by A.T. and P.M.
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Thoma, C., Toso, A., Gutbrodt, K. et al. VHL loss causes spindle misorientation and chromosome instability. Nat Cell Biol 11, 994–1001 (2009). https://doi.org/10.1038/ncb1912
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DOI: https://doi.org/10.1038/ncb1912
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