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© The Rockefeller University Press, 0021-9525/1997//385 $5.00
The Journal of Cell Biology, Volume 138, Number 2, , 1997 385-393

Article

CDK1 Inactivation Regulates Anaphase Spindle Dynamics and Cytokinesis In Vivo



Sally P. Wheatley*, Edward H. Hinchcliffe*, Michael Glotzer{ddagger}, Anthony A. Hyman{ddagger}, Greenfield Sluder*, and Yu-li Wang*

* Cell Biology Group, Worcester Foundation for Biomedical Research, Shrewsbury, Massachusetts 01545; and {ddagger} Mitotic Spindle Group, Cell Biology Program, European Molecular Biology Laboratory, 69117 Heidelberg, Germany

Through association with CDK1, cyclin B accumulation and destruction govern the G2/M/G1 transitions in eukaryotic cells. To identify CDK1 inactivation-dependent events during late mitosis, we expressed a nondestructible form of cyclin B (cyclin B{Delta}90) by microinjecting its mRNA into prometaphase normal rat kidney cells. The injection inhibited chromosome decondensation and nuclear envelope formation. Chromosome disjunction occurred normally, but anaphase-like movement persisted until the chromosomes reached the cell periphery, whereupon they often somersaulted and returned to the cell center. Injection of rhodamine-tubulin showed that this movement occurred in the absence of a central anaphase spindle. In 82% of cells cytokinesis was inhibited; the remainder split themselves into two parts in a process reminiscent of Dictyostelium cytofission. In all cells injected, F-actin and myosin II were diffusely localized with no detectable organization at the equator. Our results suggest that a primary effect of CDK1 inactivation is on spindle dynamics that regulate chromosome movement and cytokinesis. Prolonged CDK1 activity may prevent cytokinesis through inhibiting midzone microtubule formation, the behavior of proteins such as TD60, or through the phosphorylation of myosin II regulatory light chain.

Abbreviations used in this paper: NRK, normal rat kidney; NuMA, nuclear protein-associated mitotic apparatus.

This project was supported by funds from the National Institutes of Health and Human Frontier Science Program.

S.P. Wheatley would like to dedicate this work to Professor Daniel Mazia, whose insights into cell division inspired her to follow the same path in cell biology. S.P. Wheatley met Professor Mazia during a United Nations Educational, Scientific, and Cultural Organization course entitled "Dynamics of intracellular structures during the cell cycle" held in L'Ile d'Yeu, France, in the summer of 1992.

Please address all correspondence to Yu-li Wang, Cell Biology Group, Worcester Foundation for Biomedical Research, Shrewsbury, MA 01545. Tel.: (508) 845-2651; Fax: (508) 842-3915.


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