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Vol. 9, Issue 8, 2051-2068, August 1998
Department of Molecular Biology, Princeton University, Princeton,
New Jersey 08544
The roles of two kinesin-related proteins, Kip2p and Kip3p, in
microtubule function and nuclear migration were investigated. Deletion
of either gene resulted in nuclear migration defects similar to those
described for dynein and kar9 mutants. By indirect immunofluorescence, the cytoplasmic microtubules in
kip2
were consistently short or absent throughout the
cell cycle. In contrast, in kip3 strains, the
cytoplasmic microtubules were significantly longer than wild type at
telophase. Furthermore, in the kip3 cells with
nuclear positioning defects, the cytoplasmic microtubules were
misoriented and failed to extend into the bud. Localization studies
found Kip2p exclusively on cytoplasmic microtubules throughout the cell
cycle, whereas GFP-Kip3p localized to both spindle and cytoplasmic
microtubules. Genetic analysis demonstrated that the kip2 kar9 double mutants were
synthetically lethal, whereas kip3
kar9 double mutants were viable. Conversely,
kip3 dhc1 double mutants were
synthetically lethal, whereas kip2
dhc1 double mutants were viable. We suggest that the
kinesin-related proteins, Kip2p and Kip3p, function in nuclear
migration and that they do so by different mechanisms. We propose that
Kip2p stabilizes microtubules and is required as part of the
dynein-mediated pathway in nuclear migration. Furthermore, we propose
that Kip3p functions, in part, by depolymerizing microtubules and is
required for the Kar9p-dependent orientation of the cytoplasmic
microtubules.
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