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Vol. 10, Issue 10, 3081-3096, October 1999
Department of Cellular Biology, University of Georgia, Athens,
Georgia 30602-2607
We cloned two genes, KIN1 and KIN2,
encoding kinesin-II homologues from the ciliate Tetrahymena
thermophila and constructed strains lacking either
KIN1 or KIN2 or both genes. Cells with a
single disruption of either gene showed partly overlapping sets of
defects in cell growth, motility, ciliary assembly, and
thermoresistance. Deletion of both genes resulted in loss of cilia and
arrests in cytokinesis. Mutant cells were unable to assemble new cilia
or to maintain preexisting cilia. Double knockout cells were not viable
on a standard medium but could be grown on a modified medium on which
growth does not depend on phagocytosis. Double knockout cells could be
rescued by transformation with a gene encoding an epitope-tagged Kin1p.
In growing cells, epitope-tagged Kin1p preferentially accumulated in
cilia undergoing active assembly. Kin1p was also detected in the cell
body but did not show any association with the cleavage furrow. The
cell division arrests observed in kinesin-II knockout cells appear to
be induced by the loss of cilia and resulting cell paralysis.
Online version of this article contains video material
for Figure 9. Online version available at www.molbiolcell.org.
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