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© The Rockefeller University Press, 0021-9525/1998//1417 $5.00
The Journal of Cell Biology, Volume 140, Number 6, , 1998 1417-1426

Article

The Coordination of Centrosome Reproduction with Nuclear Events of the Cell Cycle in the Sea Urchin Zygote



Edward H. Hinchcliffe*, Grizzel O. Cassels{ddagger}, Conly L. Rieder{ddagger},§, and Greenfield Sluder*

* Department of Cell Biology, University of Massachusetts Medical Center, Shrewsbury, Massachusetts 01655; {ddagger} Laboratory of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York 12201-0509; and § Department of Biomedical Sciences, State University of New York, Albany, NewYork 12222

Centrosomes repeatedly reproduce in sea urchin zygotes arrested in S phase, whether cyclin-dependent kinase 1–cyclin B (Cdk1-B) activity remains at prefertilization levels or rises to mitotic values. In contrast, when zygotes are arrested in mitosis using cyclin B {Delta}-90, anaphase occurs at the normal time, yet centrosomes do not reproduce. Together, these results reveal the cell cycle stage specificity for centrosome reproduction and demonstrate that neither the level nor the cycling of Cdk1-B activity coordinate centrosome reproduction with nuclear events. In addition, the proteolytic events of the metaphase–anaphase transition do not control when centrosomes duplicate. When we block protein synthesis at first prophase, the zygotes divide and arrest before second S phase. Both blastomeres contain just two complete centrosomes, which indicates that the cytoplasmic conditions between mitosis and S phase support centrosome reproduction. However, the fact that these daughter centrosomes do not reproduce again under such supportive conditions suggests that they are lacking a component required for reproduction. The repeated reproduction of centrosomes during S phase arrest points to the existence of a necessary "licensing" event that restores this component to daughter centrosomes during S phase, preparing them to reproduce in the next cell cycle.

Abbreviations used in this paper: BrdU, 5-bromo-2'-deoxyuridine; Cdk1-B, cyclin-dependent kinase 1–cyclin B; CFSW, Ca2+-free sea water; NEB, nuclear envelope breakdown; NSW, natural sea water.

Address all correspondence to Dr. Greenfield Sluder, Department of Cell Biology, University of Massachusetts Medical Center, Shrewsbury Campus, 222 Maple Avenue, Shrewsbury, MA 01545. Tel.: (508) 842-8921, ext. 341. Fax: (508) 842-3915. E-mail: sluder{at}sci.wfbr.edu


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