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Centrosome number is controlled by a centrosome-intrinsic block to reduplication

Nature Cell Biology volume 5, pages 539–544 (2003)Cite this article

Abstract

The centrosome duplicates once in S phase. To determine whether there is a block in centrosome reduplication, we used a cell fusion assay to compare the duplication potential of unduplicated G1 centrosomes and recently duplicated G2 centrosomes. By fusing cells in different cell cycle stages, we found that G2 centrosomes were unable to reduplicate in a cellular environment that supports centrosome duplication. Furthermore, G2 cytoplasm did not inhibit centrosome duplication in fused cells, indicating that the block to reduplication is intrinsic to the centrosomes rather than the cytoplasm. To test the underlying mechanism, we created mononucleate G1 cells with two centrosomes by fusing cells with enucleated cytoplasts. Both centrosomes duplicated, indicating that the block is not controlled by centrosome:nucleus ratio. We also found that human primary cells have tight control over centrosome number during prolonged S-phase arrest and that this control is partially abrogated in transformed cells. This suggests a link between the control of centrosome duplication and maintenance of genomic stability.

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Figure 1: Cell cycle synchronization of fibroblast cells.
Figure 2: DNA replication and centrosome number in fused cells.
Figure 3: Centrosome duplication in G1–S, G2–S and G1–G2 cell fusions.
Figure 4: Centrosome duplication in G1–G1 cytoplast–cell fusions.
Figure 5: Centrosome reduplication during S-phase arrest.

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Acknowledgements

We thank J. Ford for cell lines, J. Salisbury for the anti-centrin antibody, and G.-W. Fang and P. Jackson for comments on the manuscript. This work was supported by a grant from the Human Frontier Science Program. C.W. was supported by a Stanford Graduate Fellowship.

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Authors and Affiliations

  1. Department of Biological Sciences, Stanford University, Stanford, 94305-5020, CA, USA

    Connie Wong & Tim Stearns

  2. Department of Genetics, Stanford University Medical School, Stanford, 94305-5020, CA, USA

    Tim Stearns

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  1. Connie Wong
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  2. Tim Stearns
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Correspondence to Tim Stearns.

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Wong, C., Stearns, T. Centrosome number is controlled by a centrosome-intrinsic block to reduplication. Nat Cell Biol 5, 539–544 (2003). https://doi.org/10.1038/ncb993

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