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© The Rockefeller University Press, 0021-9525/1997//1023 $5.00
The Journal of Cell Biology, Volume 138, Number 5, , 1997 1023-1040

Article

Kinesin-related KIP3 of Saccharomyces cerevisiae Is Required for a Distinct Step in Nuclear Migration



Todd M. DeZwaan*, Eric Ellingson{ddagger}, David Pellman{ddagger}, and David M. Roof*

* Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6085; and {ddagger} Departments of Pediatric Oncology, The Dana-Farber Cancer Institute, and Pediatric Hematology, The Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Spindle orientation and nuclear migration are crucial events in cell growth and differentiation of many eukaryotes. Here we show that KIP3, the sixth and final kinesin-related gene in Saccharomyces cerevisiae, is required for migration of the nucleus to the bud site in preparation for mitosis. The position of the nucleus in the cell and the orientation of the mitotic spindle was examined by microscopy of fixed cells and by time-lapse microscopy of individual live cells. Mutations in KIP3 and in the dynein heavy chain gene defined two distinct phases of nuclear migration: a KIP3-dependent movement of the nucleus toward the incipient bud site and a dynein-dependent translocation of the nucleus through the bud neck during anaphase. Loss of KIP3 function disrupts the unidirectional movement of the nucleus toward the bud and mitotic spindle orientation, causing large oscillations in nuclear position. The oscillatory motions sometimes brought the nucleus in close proximity to the bud neck, possibly accounting for the viability of a kip3 null mutant. The kip3 null mutant exhibits normal translocation of the nucleus through the neck and normal spindle pole separation kinetics during anaphase. Simultaneous loss of KIP3 and kinesin-related KAR3 function, or of KIP3 and dynein function, is lethal but does not block any additional detectable movement. This suggests that the lethality is due to the combination of sequential and possibly overlapping defects. Epitope-tagged Kip3p localizes to astral and central spindle microtubules and is also present throughout the cytoplasm and nucleus.

Abbreviations used in this paper: 5-FOA, 5-fluoroorotic acid; DAPI, 4,6-diamidino-2-phenylindole; DIC, differential interference contrast; GFP, green fluorescent protein; ts, temperature-sensitive.

Address all correspondence to David M. Roof, Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6085. Tel.: (215) 573-3636. Fax: (215) 573-5851. E-mail:roof{at}mail.med.upenn.edu


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