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:

SCFCyclin F controls centrosome homeostasis and mitotic fidelity through CP110 degradation

Abstract

Generally, F-box proteins are the substrate recognition subunits of SCF (Skp1–Cul1–F-box protein) ubiquitin ligase complexes, which mediate the timely proteolysis of important eukaryotic regulatory proteins1,2. Mammalian genomes encode roughly 70 F-box proteins, but only a handful have established functions3,4. The F-box protein family obtained its name from Cyclin F (also called Fbxo1), in which the F-box motif (the 40-amino-acid domain required for binding to Skp1) was first described5. Cyclin F, which is encoded by an essential gene, also contains a cyclin box domain, but in contrast to most cyclins, it does not bind or activate any cyclin-dependent kinases (CDKs)5,6,7. However, like other cyclins, Cyclin F oscillates during the cell cycle, with protein levels peaking in G2. Despite its essential nature and status as the founding member of the F-box protein family, Cyclin F remains an orphan protein, whose functions are unknown. Starting from an unbiased screen, we identified CP110, a protein that is essential for centrosome duplication, as an interactor and substrate of Cyclin F. Using a mode of substrate binding distinct from other F-box protein–substrate pairs, CP110 and Cyclin F physically associate on the centrioles during the G2 phase of the cell cycle, and CP110 is ubiquitylated by the SCFCyclin F ubiquitin ligase complex, leading to its degradation. siRNA-mediated depletion of Cyclin F in G2 induces centrosomal and mitotic abnormalities, such as multipolar spindles and asymmetric, bipolar spindles with lagging chromosomes. These phenotypes were reverted by co-silencing CP110 and were recapitulated by expressing a stable mutant of CP110 that cannot bind Cyclin F. Finally, expression of a stable CP110 mutant in cultured cells also promotes the formation of micronuclei, a hallmark of chromosome instability. We propose that SCFCyclin F-mediated degradation of CP110 is required for the fidelity of mitosis and genome integrity.

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: Cyclin F and CP110 interact and colocalize to the centrosomes.
Figure 2: CP110 is targeted for ubiquitylation and degradation by SCF Cyclin F during the G2 phase of the cell cycle.
Figure 3: Cyclin F silencing induces centrosome and mitotic aberrations.
Figure 4: The failure to degrade CP110 causes centrosome and mitotic defects.

Similar content being viewed by others

References

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

    Article  CAS  Google Scholar 

  2. Petroski, M. D. & Deshaies, R. J. Function and regulation of cullin-RING ubiquitin ligases. Nature Rev. Mol. Cell Biol. 6, 9–20 (2005)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  4. Skaar, J. R., D'Angiolella, V., Pagan, J. K. & Pagano, M. SnapShot: F box proteins II. Cell 137, 1358–1358 (2009)

    Article  Google Scholar 

  5. Bai, C. et al. Skp1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box. Cell 86, 263–274 (1996)

    Article  CAS  Google Scholar 

  6. Fung, T. K., Siu, W. Y., Yam, C. H., Lau, A. & Poon, R. Y. Cyclin F is degraded during G2-M by mechanisms fundamentally different from other cyclins. J. Biol. Chem. 277, 35140–35149 (2002)

    Article  CAS  Google Scholar 

  7. Tetzlaff, M. T. et al. Cyclin F disruption compromises placental development and affects normal cell cycle execution. Mol. Cell. Biol. 24, 2487–2498 (2004)

    Article  CAS  Google Scholar 

  8. Florens, L. & Washburn, M. P. Proteomic analysis by multidimensional protein identification technology. Methods Mol. Biol. 328, 159–175 (2006)

    CAS  PubMed  Google Scholar 

  9. Bai, C., Richman, R. & Elledge, S. J. Human cyclin F. EMBO J. 13, 6087–6098 (1994)

    Article  CAS  Google Scholar 

  10. Chen, Z., Indjeian, V. B., McManus, M., Wang, L. & Dynlacht, B. D. CP110, a cell cycle-dependent CDK substrate, regulates centrosome duplication in human cells. Dev. Cell 3, 339–350 (2002)

    Article  CAS  Google Scholar 

  11. Kleylein-Sohn, J. et al. Plk4-induced centriole biogenesis in human cells. Dev. Cell 13, 190–202 (2007)

    Article  CAS  Google Scholar 

  12. Dobbelaere, J. et al. A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila. PLoS Biol. 6, e224 (2008)

    Article  Google Scholar 

  13. Spektor, A., Tsang, W. Y., Khoo, D. & Dynlacht, B. D. Cep97 and CP110 suppress a cilia assembly program. Cell 130, 678–690 (2007)

    Article  CAS  Google Scholar 

  14. Kohlmaier, G. et al. Overly long centrioles and defective cell division upon excess of the SAS-4-related protein CPAP. Curr. Biol. 19, 1012–1018 (2009)

    Article  CAS  Google Scholar 

  15. Schulman, B. A., Lindstrom, D. L. & Harlow, E. Substrate recruitment to cyclin-dependent kinase 2 by a multipurpose docking site on cyclin A. Proc. Natl Acad. Sci. USA 95, 10453–10458 (1998)

    Article  ADS  CAS  Google Scholar 

  16. Balczon, R. et al. Dissociation of centrosome replication events from cycles of DNA synthesis and mitotic division in hydroxyurea-arrested Chinese hamster ovary cells. J. Cell Biol. 130, 105–115 (1995)

    Article  CAS  Google Scholar 

  17. Quintyne, N. J., Reing, J. E., Hoffelder, D. R., Gollin, S. M. & Saunders, W. S. Spindle multipolarity is prevented by centrosomal clustering. Science 307, 127–129 (2005)

    Article  ADS  CAS  Google Scholar 

  18. Nigg, E. A. Origins and consequences of centrosome aberrations in human cancers. Int. J. Cancer 119, 2717–2723 (2006)

    Article  CAS  Google Scholar 

  19. Bettencourt-Dias, M. & Glover, D. M. Centrosome biogenesis and function: centrosomics brings new understanding. Nature Rev. Mol. Cell Biol. 8, 451–463 (2007)

    Article  CAS  Google Scholar 

  20. Ganem, N. J., Godinho, S. A. & Pellman, D. A mechanism linking extra centrosomes to chromosomal instability. Nature 460, 278–282 (2009)

    Article  ADS  CAS  Google Scholar 

  21. Spruck, C. H., Won, K. A. & Reed, S. I. Deregulated cyclin E induces chromosome instability. Nature 401, 297–300 (1999)

    Article  ADS  CAS  Google Scholar 

  22. Loncarek, J. & Khodjakov, A. Ab ovo or de novo? Mechanisms of centriole duplication. Mol. Cells 27, 135–142 (2009)

    Article  CAS  Google Scholar 

  23. Guardavaccaro, D. et al. SCFßTrcp-mediated degradation of REST supports chromosomal stability by inducing the mitotic checkpoint. Nature 452, 365–369 (2008)

    Article  ADS  CAS  Google Scholar 

  24. Bassermann, F. et al. The Cdc14B-Cdh1-Plk1 axis controls the G2 DNA-damage-response checkpoint. Cell 134, 256–267 (2008)

    Article  CAS  Google Scholar 

  25. Bashir, T., Dorrello, N. V., Amador, V., Guardavaccaro, D. & Pagano, M. Control of the SCF(Skp2-Cks1) ubiquitin ligase by the APC/C(Cdh1) ubiquitin ligase. Nature 428, 190–193 (2004)

    Article  ADS  CAS  Google Scholar 

  26. Washburn, M. P., Wolters, D. & Yates, J. R. III. Large-scale analysis of the yeast proteome by multidimensional protein identification technology. Nature Biotechnol. 19, 242–247 (2001)

    Article  CAS  Google Scholar 

  27. McDonald, W. H. et al. Comparison of three directly coupled HPLC/MS, 2-phase Mud PIT, and 3-phase Mud PIT. Int. J. Mass Spectrom. 219, 245–251 (2002)

    Article  CAS  Google Scholar 

  28. Eng, J., McCormack, A. L. & Yates, J. R. III. An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J. Amer. Mass Spectrom. 5, 976–989 (1994)

    Article  CAS  Google Scholar 

  29. Tabb, D. L., McDonald, W. H. & Yates, J. R. III. DTASelect and Contrast: tools for assembling and comparing protein identifications from shotgun proteomics. J. Proteome Res. 1, 21–26 (2002)

    Article  CAS  Google Scholar 

  30. Florens, L. et al. Analyzing chromatin remodeling complexes using shotgun proteomics and normalized spectral abundance factors. Methods 40, 303–311 (2006)

    Article  CAS  Google Scholar 

  31. Paoletti, A. C. et al. Quantitative proteomic analysis of distinct mammalian Mediator complexes using normalized spectral abundance factors. Proc. Natl Acad. Sci. USA 103, 18928–18933 (2006)

    Article  ADS  CAS  Google Scholar 

  32. Zybailov, B. et al. Statistical analysis of membrane proteome expression changes in Saccharomyces cerevisiae. J. Proteome Res. 5, 2339–2347 (2006)

    Article  CAS  Google Scholar 

  33. Pagano, M., Pepperkok, R., Verde, F., Ansorge, W. & Draetta, G. Cyclin A is required at two points in the human cell cycle. EMBO J. 11, 761–771 (1992)

    Article  Google Scholar 

  34. Carrano, A. C. & Pagano, M. Role of the F-box protein Skp2 in adhesion-dependent cell cycle progression. J. Cell Biol. 153, 1381–1389 (2001)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. Elledge for Cyclin FFlox/– and Cyclin F−/− MEFs, J.R. Skaar for reading the manuscript and F.M. Forrester for technical help. M.P. is grateful to T.M. Thor for continuous support. This work was funded by fellowships from the American Italian Cancer Foundation to V.D’A. and V.D., a grant from the March of Dimes (1-FY08-372) to B.D. and grants from the National Institutes of Health (R01-GM057587, R37-CA076584 and R21-AG032560) to M.P. A.S., L.F. and M.P.W. are supported by the Stowers Institute for Medical Research. V.D’A. is a Leukemia & Lymphoma Society Fellow. M.P. is an Investigator with the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

Authors

Contributions

V.D’A. and V.D. performed and planned all experiments and helped to write the manuscript. M.P. coordinated the study, oversaw the results, and wrote the manuscript. S.V. and B.D. provided reagents, advice and assistance with the analysis of γ-tubulin and Centrin 2 foci. A.S., L.F. and M.P.W. performed the mass spectrometry analysis of the Cyclin F complex purified by V.D’A. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Michele Pagano.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-11 with legends and Supplementary Table 1. (PDF 3012 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

D’Angiolella, V., Donato, V., Vijayakumar, S. et al. SCFCyclin F controls centrosome homeostasis and mitotic fidelity through CP110 degradation. Nature 466, 138–142 (2010). https://doi.org/10.1038/nature09140

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature09140

This article is cited by

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.

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