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Vol. 11, Issue 11, 3689-3702, November 2000
and
*Department of Molecular Biology, The Scripps Research Institute,
La Jolla, California 92037; and In budding yeast, spindle polarity relies on a precise temporal
program of cytoplasmic microtubule-cortex interactions throughout spindle assembly. Loss of Clb5-dependent kinase activity under conditions of attenuated Cdc28 function disrupts this program, resulting in diploid-specific lethality. Here we show that polarity loss is tolerated by haploids due to a more prominent contribution of
microtubule-neck interactions to spindle orientation inherent to
haploids. These differences are mediated by the relative partition of
Bud6 between the bud tip and bud neck, distinguishing haploids from
diploids. Bud6 localizes initially to the bud tip and accumulates at
the neck concomitant with spindle assembly. bud6Department of Biology,
University of North Carolina, Chapel Hill, North Carolina 27599
mutant phenotypes are consistent with Bud6's role as a cortical cue
for cytoplasmic microtubule capture. Moreover, mutations that affect
Bud6 localization and partitioning disrupt the sequential program of
microtubule-cortex interactions accordingly. These data support a
model whereby Bud6 sequentially cues microtubule capture events at the
bud tip followed by capture events at the bud neck, necessary for
correct spindle morphogenesis and polarity.
Corresponding author. E-mail address:
sreed{at}scripps.edu.
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