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Fluorescence In Vivo Endomicroscopy
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Published online 18 October 2004. doi:10.1083/jcb.200407167
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 2, 231-244
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Article

 Differential contribution of Bud6p and Kar9p to microtubule capture and spindle orientation in S. cerevisiae

Stephen M. Huisman1, Olivia A.M. Bales1, Marie Bertrand1, Monique F.M.A. Smeets1, Steven I. Reed2, and Marisa Segal1

1 Department of Genetics, University of Cambridge, Cambridge, CB2 3EH UK
2 Department of Molecular Biology, MB7 The Scripps Research Institute, La Jolla, CA 92037

Correspondence to Marisa Segal: ms433{at}cam.ac.uk

In Saccharomyces cerevisiae, spindle orientation is controlled by a temporal and spatial program of microtubule (MT)–cortex interactions. This program requires Bud6p/Aip3p to direct the old pole to the bud and confine the new pole to the mother cell. Bud6p function has been linked to Kar9p, a protein guiding MTs along actin cables. Here, we show that Kar9p does not mediate Bud6p functions in spindle orientation. Based on live microscopy analysis, kar9{Delta} cells maintained Bud6p-dependent MT capture. Conversely, bud6{Delta} cells supported Kar9p-associated MT delivery to the bud. Moreover, additive phenotypes in bud6{Delta} kar9{Delta} or bud6{Delta} dyn1{Delta} mutants underscored the separate contributions of Bud6p, Kar9p, and dynein to spindle positioning. Finally, tub2C354S, a mutation decreasing MT dynamics, suppressed a kar9{Delta} mutation in a BUD6-dependent manner. Thus, Kar9p-independent capture at Bud6p sites can effect spindle orientation provided MT turnover is reduced. Together, these results demonstrate Bud6p function in MT capture at the cell cortex, independent of Kar9p-mediated MT delivery along actin cables.

Abbreviations used in this paper: DIC, differential interference contrast; MT, microtubule; SPB, spindle pole body; SPBd, daughter-bound SPB; SPBm, mother-bound SPB.


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