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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ballas, N. & Mandel, G. The many faces of REST oversee epigenetic programming of neuronal genes. Curr. Opin. Neurobiol. 15, 500–506 (2005)
Ooi, L. & Wood, I. C. Chromatin crosstalk in development and disease: lessons from REST. Nature Rev. Genet. 8, 544–554 (2007)
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)
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)
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)
Lawinger, P. et al. The neuronal repressor REST/NRSF is an essential regulator in medulloblastoma cells. Nature Med. 6, 826–831 (2000)
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)
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)
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)
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)
Westbrook, T. F. et al. A genetic screen for candidate tumor suppressors identifies REST. Cell 121, 837–848 (2005)
Guardavaccaro, D. & Pagano, M. Stabilizers and destabilizers controlling cell cycle oscillators. Mol. Cell 22, 1–4 (2006)
Jin, J. et al. Systematic analysis and nomenclature of mammalian F-box proteins. Genes Dev. 18, 2573–2580 (2004)
Dorrello, N. V. et al. S6K1- and βTRCP-mediated degradation of PDCD4 promotes protein translation and cell growth. Science 314, 467–471 (2006)
Peschiaroli, A. et al. SCFβTrCP-mediated degradation of Claspin regulates recovery from the DNA replication checkpoint response. Mol. Cell 23, 319–329 (2006)
Cardozo, T. & Pagano, M. The SCF ubiquitin ligase: insights into a molecular machine. Nature Rev. Mol. Cell Biol. 5, 739–751 (2004)
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)
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)
Busino, L. et al. Degradation of Cdc25A by β-TrCP during S phase and in response to DNA damage. Nature 426, 87–91 (2003)
Watanabe, N. et al. Ubiquitination of somatic Wee1 by SCFβTrcp is required for mitosis. Proc. Natl Acad. Sci. USA 101, 4419–4424 (2004)
Nasmyth, K. How do so few control so many? Cell 120, 739–746 (2005)
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)
Michel, L. S. et al. MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells. Nature 409, 355–359 (2001)
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)
Majumder, S. REST in good times and bad: roles in tumor suppressor and oncogenic activities. Cell Cycle 5, 1929–1935 (2006)
Pellman, D. Aneuploidy and cancer. Nature 446, 38–39 (2007)
Hernando, E. et al. Rb inactivation promotes genomic instability by uncoupling cell cycle progression from mitotic control. Nature 430, 797–802 (2004)
Kudo, Y. et al. Role of F-box protein βTrcp1 in mammary gland development and tumorigenesis. Mol. Cell. Biol. 24, 8184–8194 (2004)
Busino, L. et al. SCFFbxl3 controls the oscillation of the circadian clock by directing the degradation of cryptochrome proteins. Science 316, 900–904 (2007)
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)
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)
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)
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.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Supplementary Figures
The file contains Supplementary Figures S1-S12 with Legends. (PDF 509 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature06641
This article is cited by
-
Ubiquitin signaling in cell cycle control and tumorigenesis
Cell Death & Differentiation (2021)
-
A complex of the ubiquitin ligase TRIM32 and the deubiquitinase USP7 balances the level of c-Myc ubiquitination and thereby determines neural stem cell fate specification
Cell Death & Differentiation (2019)
-
Using proteomics to identify ubiquitin ligase–substrate pairs: how novel methods may unveil therapeutic targets for neurodegenerative diseases
Cellular and Molecular Life Sciences (2019)
-
The deubiquitylase USP15 regulates topoisomerase II alpha to maintain genome integrity
Oncogene (2018)
-
Transcriptional downregulation of miR-133b by REST promotes prostate cancer metastasis to bone via activating TGF-β signaling
Cell Death & Disease (2018)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.