Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

VHL loss causes spindle misorientation and chromosome instability

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.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: pVHL localizes to microtubules in mitosis and loss of pVHL leads to spindle misorientation.
Figure 2: pVHL stabilizes astral microtubules.
Figure 3: Loss of pVHL leads to reduced Mad2 levels and aneuploidy.
Figure 4: Depletion of pVHL in cells with a weakened spindle checkpoint leads to enhanced aneuploidy due to chromosome segregation defects.
Figure 5: Mad2 is crucial to protect cells from aneuploidy.

Similar content being viewed by others

References

  1. Frew, I. J. et al. pVHL and PTEN tumour suppressor proteins cooperatively suppress kidney cyst formation. EMBO J. 27, 1747–1757 (2008).

    Article  CAS  Google Scholar 

  2. Thoma, C. R. et al. pVHL and GSK3beta are components of a primary cilium-maintenance signalling network. Nature Cell Biol. 9, 588–595 (2007).

    Article  CAS  Google Scholar 

  3. McAinsh, A. D., Meraldi, P., Draviam, V. M., Toso, A. & Sorger, P. K. The human kinetochore proteins Nnf1R and Mcm21R are required for accurate chromosome segregation. EMBO J. 25, 4033–4049 (2006).

    Article  CAS  Google Scholar 

  4. Toyoshima, F., Matsumura, S., Morimoto, H., Mitsushima, M. & Nishida, E. PtdIns(3, 4, 5)P3 regulates spindle orientation in adherent cells. Dev. Cell 13, 796–811 (2007).

    Article  CAS  Google Scholar 

  5. Toyoshima, F. & Nishida, E. Integrin-mediated adhesion orients the spindle parallel to the substratum in an EB1- and myosin X-dependent manner. EMBO J. 26, 1487–1498 (2007).

    Article  CAS  Google Scholar 

  6. Lolkema, M. P. et al. The von Hippel-Lindau tumour suppressor interacts with microtubules through kinesin-2. FEBS Lett. 581, 4571–4576 (2007).

    Article  CAS  Google Scholar 

  7. Hergovich, A., Lisztwan, J., Barry, R., Ballschmieter, P. & Krek, W. Regulation of microtubule stability by the von Hippel-Lindau tumour suppressor protein pVHL. Nature Cell Biol. 5, 64–70 (2003).

    Article  CAS  Google Scholar 

  8. Li, L. et al. Hypoxia-inducible factor linked to differential kidney cancer risk seen with type 2A and type 2B VHL mutations. Mol. Cell Biol. 27, 5381–5392 (2007).

    Article  CAS  Google Scholar 

  9. Rieder, C. L., Cole, R. W., Khodjakov, A. & Sluder, G. The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores. J. Cell Biol. 130, 941–948 (1995).

    Article  CAS  Google Scholar 

  10. Musacchio, A. & Salmon, E. D. The spindle-assembly checkpoint in space and time. Nature Rev. Mol. Cell Biol. 8, 379–393 (2007).

    Article  CAS  Google Scholar 

  11. Weaver, B. A., Silk, A. D., Montagna, C., Verdier-Pinard, P. & Cleveland, D. W. Aneuploidy acts both oncogenically and as a tumor suppressor. Cancer Cell 11, 25–36 (2007).

    Article  CAS  Google Scholar 

  12. Tanudji, M. et al. Gene silencing of CENP-E by small interfering RNA in HeLa cells leads to missegregation of chromosomes after a mitotic delay. Mol. Biol. Cell 15, 3771–3781 (2004).

    Article  CAS  Google Scholar 

  13. Michel, L. S. et al. MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells. Nature 409, 355–359 (2001).

    Article  CAS  Google Scholar 

  14. Abou-Rebyeh, H., Borgmann, V., Nagel, R. & Al-Abadi, H. DNA ploidy is a valuable predictor for prognosis of patients with resected renal cell carcinoma. Cancer 92, 2280–2285 (2001).

    Article  CAS  Google Scholar 

  15. Pepe, S. et al. Nuclear DNA content-derived parameters correlated with heterogeneous expression of p53 and bcl-2 proteins in clear cell renal carcinomas. Cancer 89, 1065–1075 (2000).

    Article  CAS  Google Scholar 

  16. Fischer, E. et al. Defective planar cell polarity in polycystic kidney disease. Nature Genet. 38, 21–23 (2006).

    Article  CAS  Google Scholar 

  17. Solomon, D. & Schwartz, A. Renal pathology in von Hippel-Lindau disease. Hum. Pathol. 19, 1072–1079 (1988).

    Article  CAS  Google Scholar 

  18. Mandriota, S. J. et al. HIF activation identifies early lesions in VHL kidneys: evidence for site-specific tumor suppressor function in the nephron. Cancer Cell 1, 459–468 (2002).

    Article  CAS  Google Scholar 

  19. Krek, W. & Nigg, E. A. Mutations of p34cdc2 phosphorylation sites induce premature mitotic events in HeLa cells: evidence for a double block to p34cdc2 kinase activation in vertebrates. EMBO J. 10, 3331–3341 (1991).

    Article  CAS  Google Scholar 

  20. De Antoni, A. et al. The Mad1/Mad2 complex as a template for Mad2 activation in the spindle assembly checkpoint. Curr. Biol. 15, 214–225 (2005).

    Article  CAS  Google Scholar 

  21. Frew, I. J. et al. Combined VHLH and PTEN mutation causes genital tract cystadenoma and squamous metaplasia. Mol. Cell Biol. 28, 4536–4548 (2008).

    Article  CAS  Google Scholar 

  22. McClelland, S. E. et al. The CENP-A NAC/CAD kinetochore complex controls chromosome congression and spindle bipolarity. EMBO J. 26, 5033–5047 (2007).

    Article  CAS  Google Scholar 

  23. Meraldi, P., Draviam, V. M. & Sorger, P. K. Timing and checkpoints in the regulation of mitotic progression. Dev. Cell 7, 45–60 (2004).

    Article  CAS  Google Scholar 

  24. Salinas, S. et al. Human spastin has multiple microtubule-related functions. J. Neurochem. 95, 1411–1420 (2005).

    Article  CAS  Google Scholar 

  25. Mertz, K. D. et al. Association of cytokeratin 7 and 19 expression with genomic stability and favorable prognosis in clear cell renal cell cancer. Int. J. Cancer 123, 569–576 (2008).

    Article  CAS  Google Scholar 

  26. Staller, P. et al. Chemokine receptor CXCR4 downregulated by von Hippel-Lindau tumour suppressor pVHL. Nature 425, 307–311 (2003).

    Article  CAS  Google Scholar 

  27. Dalquen, P. et al. DNA aneuploidy, S-phase fraction, nuclear p53 positivity, and survival in non-small-cell lung carcinoma. Virchows Arch. 431, 173–179 (1997).

    Article  CAS  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

Authors

Contributions

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.

Corresponding authors

Correspondence to Patrick Meraldi or Wilhelm Krek.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 1056 kb)

Supplementary Information

Supplementary Video S1 (MOV 915 kb)

Supplementary Information

Supplementary Video S2 (MOV 2112 kb)

Supplementary Information

Supplementary Video S3 (MOV 2642 kb)

Supplementary Information

Supplementary Video S4 (MOV 546 kb)

Supplementary Information

Supplementary Video S5 (MOV 1389 kb)

Supplementary Information

Supplementary Video S6 (MOV 480 kb)

Supplementary Information

Supplementary Video S7 (MOV 590 kb)

Supplementary Information

Supplementary Video S8 (MOV 7013 kb)

Supplementary Information

Supplementary Video S9 (MOV 1322 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1912

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing