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Published online
doi:10.1083/jcb.200710164
The Journal of Cell Biology, Vol. 180, No. 1, 91-100
The Rockefeller University Press, 0021-9525 $30.00
© Gardner et al.
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Article

The microtubule-based motor Kar3 and plus end–binding protein Bim1 provide structural support for the anaphase spindle



Melissa K. Gardner1, Julian Haase2, Karthikeyan Mythreye2, Jeffrey N. Molk2, MaryBeth Anderson2, Ajit P. Joglekar2, Eileen T. O'Toole3, Mark Winey3, E.D. Salmon2, David J. Odde1, and Kerry Bloom2

1 Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN 55455
2 Department of Biology, University of North Carolina, Chapel Hill, NC 27599
3 Molecular, Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309

Correspondence to Kerry Bloom: kerry_bloom{at}unc.edu

In budding yeast, the mitotic spindle is comprised of 32 kinetochore microtubules (kMTs) and ~8 interpolar MTs (ipMTs). Upon anaphase onset, kMTs shorten to the pole, whereas ipMTs increase in length. Overlapping MTs are responsible for the maintenance of spindle integrity during anaphase. To dissect the requirements for anaphase spindle stability, we introduced a conditionally functional dicentric chromosome into yeast. When centromeres from the same sister chromatid attach to opposite poles, anaphase spindle elongation is delayed and a DNA breakage-fusion-bridge cycle ensues that is dependent on DNA repair proteins. We find that cell survival after dicentric chromosome activation requires the MT-binding proteins Kar3p, Bim1p, and Ase1p. In their absence, anaphase spindles are prone to collapse and buckle in the presence of a dicentric chromosome. Our analysis reveals the importance of Bim1p in maintaining a stable ipMT overlap zone by promoting polymerization of ipMTs during anaphase, whereas Kar3p contributes to spindle stability by cross-linking spindle MTs.

Abbreviations used in this paper: Chr III, chromosome III; ipMT, interpolar MT; kMT; kinetochore MT; MT, microtubule; SPB, spindle pole body.


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