Nonredundant functions of Kinesin-13s during meiotic spindle assembly.
Spatiotemporal control of microtubule depolymerization during cell division underlies the construction and dynamics of mitotic and meiotic spindles. Owing to their potent ability to disassemble microtubules, Kinesin-13s constitute an important class of microtubule destabilizing factors. Unfertilized Xenopus eggs, similar to other metazoan cells, contain the prototypical Kinesin-13 MCAK as well as a second family member, XKIF2. Here, we compare the roles of MCAK and XKIF2 during spindle assembly in Xenopus extracts. We find that although MCAK and XKIF2 have similar localization and biochemical properties, XKIF2 is not required for spindle assembly and, further, cannot substitute for MCAK. Altering dosage of the two kinesins demonstrates that spindle length is exquisitely sensitive to MCAK concentration but not XKIF2 concentration. Finally, we demonstrate that the rate of poleward microtubule flux in Xenopus-extract spindles is unaffected by XKIF2 depletion and is only modestly sensitive to reduction of MCAK action. We suggest that, in contrast to models proposed for mammalian somatic cell and embryonic Drosophila spindles, Kinesin-13s do not play a central role in poleward flux by depolymerizing minus ends. Rather, MCAK, but not XKIF2, plays a central role in regulating dynamic instability of plus ends and controls spindle length by that mechanism.
Ohi
Ryoma
R
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA. ryoma_ohi@hms.harvard.edu <ryoma_ohi@hms.harvard.edu>
Burbank
Kendra
K
Liu
Qing
Q
Mitchison
Timothy J
TJ
D000818:Animals; D002457:Cell Extracts; D016547:Kinesins / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D008954:Models, Biological; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D014981:Xenopus; D029867:Xenopus Proteins / Q000032:analysis / Q000737:chemistry / Q000502:physiology*
D002457:Cell Extracts; D029867:Xenopus Proteins; C509242:mitotic centromere-associated kinesin, Xenopus; C522444:KIF2 protein, Xenopus; D016547:Kinesins