We have identified a novel centromere-associated gene product from Saccharomyces cerevisiae that plays a role in spindle assembly and stability. Strains with a deletion of SLK19 (synthetic lethal Kar3p gene) exhibit abnormally short mitotic spindles, increased numbers of astral microtubules, and require the presence of the kinesin motor Kar3p for viability. When cells are deprived of both Slk19p and Kar3p, rapid spindle breakdown and mitotic arrest is observed. A functional fusion of Slk19p to green fluorescent protein (GFP) localizes to kinetochores and, during anaphase, to the spindle midzone, whereas Kar3p-GFP was found at the nuclear side of the spindle pole body. Thus, these proteins seem to play overlapping roles in stabilizing spindle structure while acting from opposite ends of the microtubules.
Slk19p Is a Centromere Protein That Functions to Stabilize Mitotic Spindles
Xuemei Zeng and Jason A. Kahana contributed equally to the manuscript.
This manuscript was made possible by funding support from the American Cancer Society award CB-171 to W.S. Saunders, National Institutes of Health award RR0592 to J.R. McIntosh, and a Barr Investigator award to P.A. Silver.
Abbreviations used in this paper: 5-FOA, 5-fluororotic acid; ChIP, chromatin immunoprecipitation assay; DAPI, 4′,6-diamidino-2-phenylindole; GFP, green fluorescent protein; HU, hydroxyurea; k-Mts, kinetochore microtubules; Mts, microtubules; ORF, open reading frame; SLK19, synthetic lethal Kar3p gene; SPB, spindle pole body; ts, temperature sensitive.
Xuemei Zeng, Jason A. Kahana, Pamela A. Silver, Mary K. Morphew, J. Richard McIntosh, Ian T. Fitch, John Carbon, William S. Saunders; Slk19p Is a Centromere Protein That Functions to Stabilize Mitotic Spindles. J Cell Biol 26 July 1999; 146 (2): 415–425. doi: https://doi.org/10.1083/jcb.146.2.415
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