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

Nature volume 452, pages 365–369 (2008)Cite this article

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.

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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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Authors and Affiliations

  1. Department of Pathology, and,

    Daniele Guardavaccaro, David Frescas, N. Valerio Dorrello, Angelo Peschiaroli, Eva Hernando & Michele Pagano

  2. Department of Pharmacology, NYU Cancer Institute, New York University School of Medicine, 550 First Avenue, MSB 599, New York, New York 10016, USA,

    Timothy Cardozo

  3. Department of Cancer Genetics, The MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77030, USA,

    Asha S. Multani & Sandy Chang

  4. Institute for Cancer Genetics, Columbia University New York, New York 10032, USA

    Anna Lasorella & Antonio Iavarone

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  1. Daniele Guardavaccaro
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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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