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Control of chromosome stability by the β-TrCP–REST–Mad2 axis

Abstract

REST/NRSF (repressor-element-1-silencing transcription factor/neuron-restrictive silencing factor) negatively regulates the transcription of genes containing RE1 sites1,2. REST is expressed in non-neuronal cells and stem/progenitor neuronal cells, in which it inhibits the expression of neuron-specific genes. Overexpression of REST is frequently found in human medulloblastomas and neuroblastomas3,4,5,6,7, in which it is thought to maintain the stem character of tumour cells. Neural stem cells forced to express REST and c-Myc fail to differentiate and give rise to tumours in the mouse cerebellum3. Expression of a splice variant of REST that lacks the carboxy terminus has been associated with neuronal tumours and small-cell lung carcinomas8,9,10, and a frameshift mutant (REST-FS), which is also truncated at the C terminus, has oncogenic properties11. Here we show, by using an unbiased screen, that REST is an interactor of the F-box protein β-TrCP. REST is degraded by means of the ubiquitin ligase SCFβ-TrCP during the G2 phase of the cell cycle to allow transcriptional derepression of Mad2, an essential component of the spindle assembly checkpoint. The expression in cultured cells of a stable REST mutant, which is unable to bind β-TrCP, inhibited Mad2 expression and resulted in a phenotype analogous to that observed in Mad2+/- cells. In particular, we observed defects that were consistent with faulty activation of the spindle checkpoint, such as shortened mitosis, premature sister-chromatid separation, chromosome bridges and mis-segregation in anaphase, tetraploidy, and faster mitotic slippage in the presence of a spindle inhibitor. An indistinguishable phenotype was observed by expressing the oncogenic REST-FS mutant11, which does not bind β-TrCP. Thus, SCFβ-TrCP-dependent degradation of REST during G2 permits the optimal activation of the spindle checkpoint, and consequently it is required for the fidelity of mitosis. The high levels of REST or its truncated variants found in certain human tumours may contribute to cellular transformation by promoting genomic instability.

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Figure 1: REST is targeted for degradation by SCF β-TrCP during G2.
Figure 2: Mad2 is a transcriptional target of REST.
Figure 3: Failure to degrade REST causes defects in the mitotic checkpoint.
Figure 4: Expression of a stable REST mutant or oncogenic REST-FS leads to chromosomal instability.

References

  1. Ballas, N. & Mandel, G. The many faces of REST oversee epigenetic programming of neuronal genes. Curr. Opin. Neurobiol. 15, 500–506 (2005)

    CAS  Article  Google Scholar 

  2. Ooi, L. & Wood, I. C. Chromatin crosstalk in development and disease: lessons from REST. Nature Rev. Genet. 8, 544–554 (2007)

    CAS  Article  Google Scholar 

  3. Su, X. et al. Abnormal expression of REST/NRSF and Myc in neural stem/progenitor cells causes cerebellar tumors by blocking neuronal differentiation. Mol. Cell. Biol. 26, 1666–1678 (2006)

    CAS  Article  Google Scholar 

  4. Fuller, G. N. et al. Many human medulloblastoma tumors overexpress repressor element-1 silencing transcription (REST)/neuron-restrictive silencer factor, which can be functionally countered by REST-VP16. Mol. Cancer Ther. 4, 343–349 (2005)

    CAS  PubMed  Google Scholar 

  5. Higashino, K., Narita, T., Taga, T., Ohta, S. & Takeuchi, Y. Malignant rhabdoid tumor shows a unique neural differentiation as distinct from neuroblastoma. Cancer Sci. 94, 37–42 (2003)

    CAS  Article  Google Scholar 

  6. Lawinger, P. et al. The neuronal repressor REST/NRSF is an essential regulator in medulloblastoma cells. Nature Med. 6, 826–831 (2000)

    CAS  Article  Google Scholar 

  7. Nishimura, E., Sasaki, K., Maruyama, K., Tsukada, T. & Yamaguchi, K. Decrease in neuron-restrictive silencer factor (NRSF) mRNA levels during differentiation of cultured neuroblastoma cells. Neurosci. Lett. 211, 101–104 (1996)

    CAS  Article  Google Scholar 

  8. Gurrola-Diaz, C., Lacroix, J., Dihlmann, S., Becker, C. M. & von Knebel Doeberitz, M. Reduced expression of the neuron restrictive silencer factor permits transcription of glycine receptor α1 subunit in small-cell lung cancer cells. Oncogene 22, 5636–5645 (2003)

    CAS  Article  Google Scholar 

  9. Neumann, S. B. et al. Relaxation of glycine receptor and onconeural gene transcription control in NRSF deficient small cell lung cancer cell lines. Brain Res. Mol. Brain Res. 120, 173–181 (2004)

    CAS  Article  Google Scholar 

  10. Coulson, J. M., Edgson, J. L., Woll, P. J. & Quinn, J. P. A splice variant of the neuron-restrictive silencer factor repressor is expressed in small cell lung cancer: a potential role in derepression of neuroendocrine genes and a useful clinical marker. Cancer Res. 60, 1840–1844 (2000)

    CAS  PubMed  Google Scholar 

  11. Westbrook, T. F. et al. A genetic screen for candidate tumor suppressors identifies REST. Cell 121, 837–848 (2005)

    CAS  Article  Google Scholar 

  12. Guardavaccaro, D. & Pagano, M. Stabilizers and destabilizers controlling cell cycle oscillators. Mol. Cell 22, 1–4 (2006)

    CAS  Article  Google Scholar 

  13. Jin, J. et al. Systematic analysis and nomenclature of mammalian F-box proteins. Genes Dev. 18, 2573–2580 (2004)

    CAS  Article  Google Scholar 

  14. Dorrello, N. V. et al. S6K1- and βTRCP-mediated degradation of PDCD4 promotes protein translation and cell growth. Science 314, 467–471 (2006)

    ADS  CAS  Article  Google Scholar 

  15. Peschiaroli, A. et al. SCFβTrCP-mediated degradation of Claspin regulates recovery from the DNA replication checkpoint response. Mol. Cell 23, 319–329 (2006)

    CAS  Article  Google Scholar 

  16. Cardozo, T. & Pagano, M. The SCF ubiquitin ligase: insights into a molecular machine. Nature Rev. Mol. Cell Biol. 5, 739–751 (2004)

    CAS  Article  Google Scholar 

  17. Wu, G. et al. Structure of a β-TrCP1–Skp1–β-catenin complex: destruction motif binding and lysine specificity of the SCFβ-TrCP1 ubiquitin ligase. Mol. Cell 11, 1445–1456 (2003)

    CAS  Article  Google Scholar 

  18. Guardavaccaro, D. et al. Control of meiotic and mitotic progression by the F box protein β-Trcp1 in vivo. Dev. Cell 4, 799–812 (2003)

    CAS  Article  Google Scholar 

  19. Busino, L. et al. Degradation of Cdc25A by β-TrCP during S phase and in response to DNA damage. Nature 426, 87–91 (2003)

    ADS  CAS  Article  Google Scholar 

  20. Watanabe, N. et al. Ubiquitination of somatic Wee1 by SCFβTrcp is required for mitosis. Proc. Natl Acad. Sci. USA 101, 4419–4424 (2004)

    ADS  CAS  Article  Google Scholar 

  21. Nasmyth, K. How do so few control so many? Cell 120, 739–746 (2005)

    CAS  Article  Google Scholar 

  22. Latres, E., Chiaur, J. & Pagano, M. The human F box protein β-Trcp associates with the Cul1/Skp1 complex and regulates the stability of β-catenin. Oncogene 18, 849–854 (1999)

    CAS  Article  Google Scholar 

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

    ADS  CAS  Article  Google Scholar 

  24. Ballas, N., Grunseich, C., Lu, D. D., Speh, J. C. & Mandel, G. REST and its corepressors mediate plasticity of neuronal gene chromatin throughout neurogenesis. Cell 121, 645–657 (2005)

    CAS  Article  Google Scholar 

  25. Majumder, S. REST in good times and bad: roles in tumor suppressor and oncogenic activities. Cell Cycle 5, 1929–1935 (2006)

    CAS  Article  Google Scholar 

  26. Pellman, D. Aneuploidy and cancer. Nature 446, 38–39 (2007)

    ADS  CAS  Article  Google Scholar 

  27. Hernando, E. et al. Rb inactivation promotes genomic instability by uncoupling cell cycle progression from mitotic control. Nature 430, 797–802 (2004)

    ADS  CAS  Article  Google Scholar 

  28. Kudo, Y. et al. Role of F-box protein βTrcp1 in mammary gland development and tumorigenesis. Mol. Cell. Biol. 24, 8184–8194 (2004)

    CAS  Article  Google Scholar 

  29. Busino, L. et al. SCFFbxl3 controls the oscillation of the circadian clock by directing the degradation of cryptochrome proteins. Science 316, 900–904 (2007)

    ADS  CAS  Article  Google Scholar 

  30. Amador, V., Ge, S., Santamaria, P., Guardavaccaro, D. & Pagano, M. APC/CCdc20 controls the ubiquitin-mediated degradation of p21 in prometaphase. Mol. Cell 27, 462–473 (2000)

    Article  Google Scholar 

  31. Nemethy, G. et al. Energy parameters in polypeptides. 10. Improved geometric parameters and nonbonded interactions for use in the ECEPP/3 algorithm with application to proline-containing peptides. J. Phys. Chem. 96, 6472–6484 (1992)

    CAS  Article  Google Scholar 

  32. Totrov, M. & Abagyan, R. Rapid boundary element solvation electrostatics calculations in folding simulations: successful folding of a 23-residue peptide. Biopolymers 60, 124–133 (2001)

    CAS  Article  Google Scholar 

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Acknowledgements

We thank V. D’Angiolella, S. Ge, L. Gnatovskiy, J. Staveroski and N. E. Sherman for their contributions to this work; P. Jallepalli and J. Skaar for suggestions and/or critically reading the manuscript; S. Elledge and T. Westbrook for communicating results before publication; and the T. C. Hsu Molecular Cytogenics Core. M.P. is grateful to T. M. Thor for continuous support. D.G. is grateful to R. Dolce and L. Guardavaccaro. This work was supported by an Emerald Foundation grant to D.G., a fellowship from Provincia di Benevento to D.G., American–Italian Cancer Foundation fellowships to D.G., N.V.D. and A.P., and grants from the National Institutes of Health to S.C. and M.P.

Author Contributions D.G. performed and planned all experiments (except chromosome analysis in Fig. 4b, which was performed by A.S.M. and S.C, and the β-TrCP immunopurifications, which were performed by N.V.D. and A.P.) and helped to write the manuscript. M.P. coordinated the study, oversaw the results and wrote the manuscript. D.F. contributed to time-lapse experiments. E.H. provided reagents and suggestions. T.C. developed the interaction models. A.L. and A.I. performed unpublished experiments to analyse stem cell differentiation. All authors discussed the results and commented on the manuscript.

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Correspondence to Michele Pagano.

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Guardavaccaro, D., Frescas, D., Dorrello, N. et al. Control of chromosome stability by the β-TrCP–REST–Mad2 axis. Nature 452, 365–369 (2008). https://doi.org/10.1038/nature06641

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