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Vol. 13, Issue 6, 1832-1845, June 2002
Department of Genetics, Cell Biology and Development, University of
Minnesota, Minneapolis, Minnesota 55455
CHO1 is a mammalian kinesin-like motor protein of the MKLP1
subfamily. It associates with the spindle midzone during anaphase and
concentrates to a midbody matrix during cytokinesis. CHO1 was
originally implicated in karyokinesis, but the invertebrate homologues
of CHO1 were shown to function in the midzone formation and
cytokinesis. To analyze the role of the protein in mammalian cells, we
mutated the ATP-binding site of CHO1 and expressed it in CHO cells.
Mutant protein (CHO1F') was able to interact with microtubules via
ATP-independent microtubule-binding site(s) but failed to accumulate at
the midline of the central spindle and affected the localization of
endogenous CHO1. Although the segregation of chromosomes, the bundling
of midzone microtubules, and the initiation of cytokinesis proceeded
normally in CHO1F'-expressing cells, the completion of cytokinesis was
inhibited. Daughter cells were frequently entering interphase while
connected by a microtubule-containing cytoplasmic bridge from which the
dense midbody matrix was missing. Depletion of endogenous CHO1 via
RNA-mediated interference also affected the formation of midbody matrix
in dividing cells, caused the disorganization of midzone microtubules,
and resulted in abortive cytokinesis. Thus, CHO1 may not be required
for karyokinesis, but it is essential for the proper midzone/midbody
formation and cytokinesis in mammalian cells.
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