Efficient mitosis in human cells lacking poleward microtubule flux.
Chromosome segregation relies on the dynamic properties of spindle microtubules (MTs). Poleward MT flux contributes to spindle dynamics through the disassembly of MT minus ends at spindle poles coupled to the continuous poleward transport of spindle MTs. Despite being conserved in metazoan cells, the function of flux remains controversial because flux rates differ widely in different cell types. In meiotic systems, the rate of flux nearly matches that of chromosome movement, but in mitotic systems, flux is significantly slower than chromosome movement. Here, we show that spindles in human mitotic cells depleted of the kinesin-13 proteins Kif2a and MCAK lack detectable flux and that such cells frequently fail to segregate all chromosomes appropriately at anaphase. Elimination of flux reduces poleward chromosome velocity approximately 20%, but does not hinder bipolar spindle assembly, chromosome alignment, or mitotic progression. Thus, mitosis proceeds efficiently in human cells lacking detectable poleward MT flux. These data demonstrate that in human cultured cells, kinetochores are sufficient to effectively power chromosome movement, leading us to speculate that flux is maintained in these cells to fulfill other functional roles such as error correction or kinetochore regulation.
Ganem
Neil J
NJ
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Upton
Kristi
K
Compton
Duane A
DA
D001692:Biological Transport / Q000502:physiology; D002478:Cells, Cultured; D020090:Chromosome Segregation / Q000502:physiology*; D049452:Green Fluorescent Proteins; D006801:Humans; D016547:Kinesins / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000502:physiology*; D008938:Mitosis / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D008941:Spindle Apparatus / Q000502:physiology*; D014404:Tubulin
C091489:KIF2C protein, human; D009419:Nerve Tissue Proteins; D012097:Repressor Proteins; D014404:Tubulin; D049452:Green Fluorescent Proteins; C477238:KIF2A protein, mouse; D016547:Kinesins