The chromokinesin Kid is necessary for chromosome arm orientation and oscillation, but not congression, on mitotic spindles.
Chromokinesins have been postulated to provide the polar ejection force needed for chromosome congression during mitosis. We have evaluated that possibility by monitoring chromosome movement in vertebrate-cultured cells using time-lapse differential interference contrast microscopy after microinjection with antibodies specific for the chromokinesin Kid. 17.5% of cells injected with Kid-specific antibodies have one or more chromosomes that remain closely opposed to a spindle pole and fail to enter anaphase. In contrast, 82.5% of injected cells align chromosomes in metaphase, progress to anaphase, and display chromosome velocities not significantly different from control cells. However, injected cells lack chromosome oscillations, and chromosome orientation is atypical because chromosome arms extend toward spindle poles during both congression and metaphase. Furthermore, chromosomes cluster into a mass and fail to oscillate when Kid is perturbed in cells containing monopolar spindles. These data indicate that Kid generates the polar ejection force that pushes chromosome arms away from spindle poles in vertebrate-cultured cells. This force increases the efficiency with which chromosomes make bipolar spindle attachments and regulates kinetochore activities necessary for chromosome oscillation, but is not essential for chromosome congression.
Levesque
A A
AA
Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755, USA.
Compton
D A
DA
D000911:Antibodies, Monoclonal; D016764:Cell Polarity / Q000187:drug effects; D002875:Chromosomes / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D004268:DNA-Binding Proteins / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D007700:Kinetics; D008845:Microinjections; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008938:Mitosis; D020409:Molecular Motor Proteins / Q000378:metabolism / Q000502:physiology; D009068:Movement; D009687:Nuclear Proteins / Q000378:metabolism / Q000502:physiology*; D021381:Protein Transport; D008941:Spindle Apparatus / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004268:DNA-Binding Proteins; C098764:KIF22 protein, human; D020409:Molecular Motor Proteins; D009687:Nuclear Proteins; C092349:chromokinesin; D016547:Kinesins