Skip to main content

Thank you for visiting 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.

Mechanism limiting centrosome duplication to once per cell cycle


The centrosome organizes the microtubule cytoskeleton and consists of a pair of centrioles surrounded by pericentriolar material. Cells begin the cell cycle with a single centrosome, which duplicates once before mitosis. During duplication, new centrioles grow orthogonally to existing ones and remain engaged (tightly opposed) with those centrioles until late mitosis or early G1 phase, when they become disengaged1. The relationship between centriole engagement/disengagement and centriole duplication potential is not understood, and the mechanisms that control these processes are not known. Here we show that centriole disengagement requires the protease separase2 at anaphase, and that this disengagement licences centriole duplication in the next cell cycle. We describe an in vitro system using Xenopus egg extract and purified centrioles in which both centriole disengagement and centriole growth occur. Centriole disengagement at anaphase is independent of mitotic exit and Cdk2/cyclin E activity, but requires the anaphase-promoting complex and separase. In contrast to disengagement, new centriole growth occurs in interphase, is dependent on Cdk2/cyclin E, and requires previously disengaged centrioles. This suggests that re-duplication of centrioles within a cell cycle is prevented by centriole engagement itself. We propose that the ‘once-only’ control of centrosome duplication is achieved by temporally separating licensing in anaphase from growth of new centrioles during S phase. The involvement of separase in both centriole disengagement and sister chromatid separation would prevent premature centriole disengagement before anaphase onset, which can lead to multipolar spindles and genomic instability3,4.

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

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it


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

Figure 1: Centriole disengagement activity is present in late mitosis.
Figure 2: Molecular requirements for centriole disengagement.
Figure 3: Separase is essential for centriole disengagement.
Figure 4: Centriole disengagement is required for centriole growth.


  1. Kuriyama, R. & Borisy, G. G. Centriole cycle in Chinese hamster ovary cells as determined by whole-mount electron microscopy. J. Cell Biol. 91, 814–821 (1981)

    Article  CAS  Google Scholar 

  2. Nasmyth, K., Peters, J. M. & Uhlmann, F. Splitting the chromosome: cutting the ties that bind sister chromatids. Science 288, 1379–1385 (2000)

    Article  ADS  CAS  Google Scholar 

  3. Hut, H. M. et al. Centrosomes split in the presence of impaired DNA integrity during mitosis. Mol. Biol. Cell 14, 1993–2004 (2003)

    Article  CAS  Google Scholar 

  4. McDermott, K. M. et al. p16(INK4a) prevents centrosome dysfunction and genomic instability in primary cells. PLoS Biol. 4, e51 (2006)

    Article  Google Scholar 

  5. Bahe, S., Stierhof, Y. D., Wilkinson, C. J., Leiss, F. & Nigg, E. A. Rootletin forms centriole-associated filaments and functions in centrosome cohesion. J. Cell Biol. 171, 27–33 (2005)

    Article  CAS  Google Scholar 

  6. Hinchcliffe, E. H., Li, C., Thompson, E. A., Maller, J. L. & Sluder, G. Requirement of Cdk2-cyclin E activity for repeated centrosome reproduction in Xenopus egg extracts. Science 283, 851–854 (1999)

    Article  ADS  CAS  Google Scholar 

  7. Lacey, K. R., Jackson, P. K. & Stearns, T. Cyclin-dependent kinase control of centrosome duplication. Proc. Natl Acad. Sci. USA 96, 2817–2822 (1999)

    Article  ADS  CAS  Google Scholar 

  8. Matsumoto, Y., Hayashi, K. & Nishida, E. Cyclin-dependent kinase 2 (Cdk2) is required for centrosome duplication in mammalian cells. Curr. Biol. 9, 429–432 (1999)

    Article  CAS  Google Scholar 

  9. Meraldi, P., Lukas, J., Fry, A. M., Bartek, J. & Nigg, E. A. Centrosome duplication in mammalian somatic cells requires E2F and Cdk2-cyclin A. Nature Cell Biol. 1, 88–93 (1999)

    Article  CAS  Google Scholar 

  10. Wong, C. & Stearns, T. Centrosome number is controlled by a centrosome-intrinsic block to reduplication. Nature Cell Biol. 5, 539–544 (2003)

    Article  CAS  Google Scholar 

  11. Gard, D. L., Hafezi, S., Zhang, T. & Doxsey, S. J. Centrosome duplication continues in cycloheximide-treated Xenopus blastulae in the absence of a detectable cell cycle. J. Cell Biol. 110, 2033–2042 (1990)

    Article  CAS  Google Scholar 

  12. Vidwans, S. J., Wong, M. L. & O'Farrell, P. H. Mitotic regulators govern progress through steps in the centrosome duplication cycle. J. Cell Biol. 147, 1371–1378 (1999)

    Article  CAS  Google Scholar 

  13. Mayor, T., Stierhof, Y. D., Tanaka, K., Fry, A. M. & Nigg, E. A. The centrosomal protein C-Nap1 is required for cell cycle-regulated centrosome cohesion. J. Cell Biol. 151, 837–846 (2000)

    Article  CAS  Google Scholar 

  14. Callaini, G. & Riparbelli, M. G. Centriole and centrosome cycle in the early Drosophila embryo. J. Cell Sci. 97, 539–543 (1990)

    PubMed  Google Scholar 

  15. Tsou, M. F. & Stearns, T. Controlling centrosome number: licenses and blocks. Curr. Opin. Cell Biol. 18, 74–78 (2006)

    Article  CAS  Google Scholar 

  16. Paoletti, A., Moudjou, M., Paintrand, M., Salisbury, J. L. & Bornens, M. Most of centrin in animal cells is not centrosome-associated and centrosomal centrin is confined to the distal lumen of centrioles. J. Cell Sci. 109, 3089–3102 (1996)

    CAS  PubMed  Google Scholar 

  17. Guadagno, T. M. & Newport, J. W. Cdk2 kinase is required for entry into mitosis as a positive regulator of Cdc2-cyclin B kinase activity. Cell 84, 73–82 (1996)

    Article  CAS  Google Scholar 

  18. Holloway, S. L., Glotzer, M., King, R. W. & Murray, A. W. Anaphase is initiated by proteolysis rather than by the inactivation of maturation-promoting factor. Cell 73, 1393–1402 (1993)

    Article  CAS  Google Scholar 

  19. Stemmann, O., Zou, H., Gerber, S. A., Gygi, S. P. & Kirschner, M. W. Dual inhibition of sister chromatid separation at metaphase. Cell 107, 715–726 (2001)

    Article  CAS  Google Scholar 

  20. Zou, H., McGarry, T. J., Bernal, T. & Kirschner, M. W. Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis. Science 285, 418–422 (1999)

    Article  CAS  Google Scholar 

  21. Nishitani, H. & Lygerou, Z. Control of DNA replication licensing in a cell cycle. Genes Cells 7, 523–534 (2002)

    Article  CAS  Google Scholar 

  22. Peloponese, J. M. Jr, Haller, K., Miyazato, A. & Jeang, K. T. Abnormal centrosome amplification in cells through the targeting of Ran-binding protein-1 by the human T cell leukemia virus type-1 Tax oncoprotein. Proc. Natl Acad. Sci. USA 102, 18974–18979 (2005)

    Article  ADS  CAS  Google Scholar 

  23. Liu, B. et al. Human T-lymphotropic virus type 1 oncoprotein tax promotes unscheduled degradation of Pds1p/securin and Clb2p/cyclin B1 and causes chromosomal instability. Mol. Cell. Biol. 23, 5269–5281 (2003)

    Article  CAS  Google Scholar 

  24. Nagao, K., Adachi, Y. & Yanagida, M. Separase-mediated cleavage of cohesin at interphase is required for DNA repair. Nature 430, 1044–1048 (2004)

    Article  ADS  CAS  Google Scholar 

  25. Gimenez-Abian, J. F., Diaz-Martinez, L. A., Waizenegger, I. C., Gimenez-Martin, G. & Clarke, D. J. Separase is required at multiple pre-anaphase cell cycle stages in human cells. Cell Cycle 4, 1576–1584 (2005)

    Article  CAS  Google Scholar 

  26. Wirth, K. G. et al. Separase: a universal trigger for sister chromatid disjunction but not chromosome cycle progression. J. Cell Biol. 172, 847–860 (2006)

    Article  CAS  Google Scholar 

  27. Kumada, K. et al. The selective continued linkage of centromeres from mitosis to interphase in the absence of mammalian separase. J. Cell Biol. 172, 835–846 (2006)

    Article  CAS  Google Scholar 

  28. Nigg, E. A. Centrosome aberrations: cause or consequence of cancer progression? Nature Rev. Cancer 2, 815–825 (2002)

    Article  CAS  Google Scholar 

Download references


We thank H. Zou for the non-degradable securin construct; P. Jackson for the D-box peptide and the Δ34Xic1 construct; L. Rose for comments on the manuscript; and J. Lüders for helpful discussions. This work was supported by a grant to T.S. from the National Institutes of Health. M.-F.B.T. is a fellow supported by the Damon-Runyon Cancer Research Foundation.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Tim Stearns.

Ethics declarations

Competing interests

Reprints and permissions information is available at The authors declare no competing financial interests.

Supplementary information

Supplementary Figure 1

Centriole configuration in the cell cycle. (PDF 361 kb)

Supplementary Figure 2

A model for regulation of centrosome duplication. (PDF 99 kb)

Supplementary Figure Legends

Text to accompany the above Supplementary Figures. (DOC 28 kb)

Supplementary Methods

This file contains additional details of the methods used in this study. (DOC 69 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tsou, MF., Stearns, T. Mechanism limiting centrosome duplication to once per cell cycle. Nature 442, 947–951 (2006).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


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.


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