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Originally published as MBC in Press, 10.1091/mbc.E03-04-0249 on August 7, 2003

Vol. 14, Issue 11, 4685-4694, November 2003

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Localization and Dynamics of Cdc2-Cyclin B during Meiotic Reinitiation in Starfish Oocytes

Mark Terasaki * {dagger}, Ei-ichi Okumura {ddagger}, Beth Hinkle * §, and Takeo Kishimoto {ddagger}

* Department of Physiology, University of Connecticut Health Center, Farmington, Connecticut 06032; {ddagger} Laboratory of Cell and Developmental Biology, Graduate School of Bioscience, Tokyo Institute of Technology, Nagatsuta, Midoriku, Yokohama 226-8501, Japan

Submitted April 21, 2003; Revised July 11, 2003; Accepted July 11, 2003
Monitoring Editor: Mark Solomon

The Cdc2-cyclin B kinase has a central role in regulating the onset of M phase. In starfish oocytes, Cdc2-cyclin B begins to be activated ~10 min after application of maturation hormone, followed by accumulation in the nucleus then nuclear envelope breakdown. By immunofluorescence and by expressing a green fluorescent (GFP) chimera of cyclin B, we find that cyclin B is present in aggregates in the cytoplasm of immature oocytes. The aggregates disperse at ~10 min, suggesting that the dispersal is closely related to the activation of the kinase. Using cyclin B-GFP, the dispersion begins from the region containing the centrosomes. Extractability of Cdc2-cyclin B changes with similar kinetics during maturation. Active Cdc25 phosphatase released Cdc2-cyclin B from the detergent-insoluble fraction independently of its phosphatase activity. Live cell imaging also showed that Cdc2-cyclin B begins to accumulate in the nucleus before changes in nuclear pore permeability, consistent with Cdc2-cyclin B-induced disassembly of the pores.

Online version of this article contains video materials for several figures. Online version is available at www.molbiolcell.org.

§ Present address: Miravant Medical Technologies, Goleta, CA 93117.

{dagger} Corresponding author. E-mail: terasaki{at}neuron.uchc.edu.




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