Dose-dependent effects of stable cyclin B1 on progression through mitosis in human cells.
The disassembly of the mitotic spindle and exit from mitosis require the inactivation of Cdk1. Here, we show that expression of nondegradable cyclinB1 causes dose-dependent mitotic arrest phenotypes. By monitoring chromosomes in living cells, we determined that pronounced overexpression of stable cyclinB1 entailed metaphase arrest without detectable sister chromatid separation, while moderate overexpression arrested cells in a pseudometaphase state, in which separated sister chromatids were kept at the cellular equator by a bipolar 'metaphase-like' spindle. Chromosomes that left the pseudometaphase plate became pulled back and individual kinetochores were found to be merotelically attached to both spindle poles in stable cyclinB1 arrested cells. Inactivation of the chromokinesin hKid, by RNAi or antibody microinjection, prevented the formation of stable bipolar spindles and the 'metaphase-like' alignment of chromosomes in cells expressing stable cyclinB1. These experiments show that cyclinB1 is able to maintain a bipolar spindle even after sister chromatids had become separated and suggest an important role of hKid in this process. Cells expressing low levels of nondegradable cyclinB1 progressed further in mitosis and arrested in telophase.
Wolf
Frank
F
Division of Molecular Pathophysiology, Biocenter, Innsbruck Medical University, Innsbruck, Austria.
Wandke
Cornelia
C
Isenberg
Nina
N
Geley
Stephan
S
D000705:Anaphase / Q000502:physiology; D000818:Animals; D002842:Chromatids / Q000235:genetics; D002877:Chromosomes, Human / Q000235:genetics; D019926:Cyclin B / Q000378:metabolism*; D056744:Cyclin B1; D004268:DNA-Binding Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism; D008677:Metaphase / Q000502:physiology; D051379:Mice; D008938:Mitosis / Q000502:physiology*; D009687:Nuclear Proteins / Q000378:metabolism; D010641:Phenotype; D008941:Spindle Apparatus / Q000378:metabolism; D013692:Telophase / Q000502:physiology; D014407:Tumor Cells, Cultured
C534699:CCNB1 protein, human; C534700:Ccnb1 protein, mouse; D019926:Cyclin B; D056744:Cyclin B1; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C092349:chromokinesin; D016547:Kinesins