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Published online 23 September 2002. doi:10.1083/jcb.200205102
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© The Rockefeller University Press, 0021-9525/2002/9/1171 $5.00
The Journal of Cell Biology, Volume 158, Number 7, September 30, 2002 1171-1181

Article

De novo formation of centrosomes in vertebrate cells arrested during S phase



Alexey Khodjakov1,2, Conly L. Rieder1,2, Greenfield Sluder3, Grisel Cassels1, Ody Sibon4 and Chuo-Lung Wang5,6

1 Division of Molecular Medicine, Wadsworth Center, New York State Department of Health, Albany, NY 12201
2 Department of Biomedical Sciences, State University of New York, Albany, NY 12222
3 Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01605
4 Department of Radiation and Stress Cell Biology, Medical Sciences, University of Groningen, 9713 AV Groningen, Netherlands
5 AutoQuant Imaging, Inc., Watervliet, NY 12189
6 Rensselaer Polytechnic Institute, Rensselaer, NY 12180

Address correspondence to Alexy Khodjakov, Wadsworth Center, P.O. Box 509, Albany, NY 12201-0509. Tel.: (518) 486-5339. Fax: (518) 486-4901. E-mail: khodj{at}wadsworth.org

The centrosome usually replicates in a semiconservative fashion, i.e., new centrioles form in association with preexisting "maternal" centrioles. De novo formation of centrioles has been reported for a few highly specialized cell types but it has not been seen in vertebrate somatic cells. We find that when centrosomes are completely destroyed by laser microsurgery in CHO cells arrested in S phase by hydroxyurea, new centrosomes form by de novo assembly. Formation of new centrosomes occurs in two steps: ~5–8 h after ablation, clouds of pericentriolar material (PCM) containing {gamma}-tubulin and pericentrin appear in the cell. By 24 h, centrioles have formed inside of already well-developed PCM clouds. This de novo pathway leads to the formation of a random number of centrioles (2–14 per cell). Although clouds of PCM consistently form even when microtubules are completely disassembled by nocodazole, the centrioles are not assembled under these conditions.

Key Words: centrosome; centrioles; microtubules; microtubule-organizing centers; cell cycle


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