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Vol. 11, Issue 4, 1197-1211, April 2000
Abteilung Molekulare Mikrobiologie, Biozentrum, Universität
Basel, CH-4056 Basel, Switzerland
Saccharomyces cerevisiae cnm67
cells
lack the spindle pole body (SPB) outer plaque, the main attachment site
for astral (cytoplasmic) microtubules, leading to frequent nuclear
segregation failure. We monitored dynamics of green fluorescent
protein-labeled nuclei and microtubules over several cell cycles.
Early nuclear migration steps such as nuclear positioning and spindle
orientation were slightly affected, but late phases such as rapid
oscillations and insertion of the anaphase nucleus into the bud neck
were mostly absent. Analyzes of microtubule dynamics revealed normal
behavior of the nuclear spindle but frequent detachment of astral
microtubules after SPB separation. Concomitantly, Spc72 protein, the
cytoplasmic anchor for the 
-tubulin complex, was partially lost from
the SPB region with dynamics similar to those observed for
microtubules. We postulate that in cnm67 cells
Spc72--tubulin complex-capped astral microtubules are released from
the half-bridge upon SPB separation but fail to be anchored to the
cytoplasmic side of the SPB because of the absence of an outer plaque.
However, successful nuclear segregation in cnm67
cells can still be achieved by elongation forces of spindles that were
correctly oriented before astral microtubule detachment by action of
Kip3/Kar3 motors. Interestingly, the first nuclear segregation in
newborn diploid cells never fails, even though astral microtubule
detachment occurs.
Online version of this article contains video material
for Figures 1-7. Online version available at www.molbiolcell.org.
*
Corresponding author. E-mail address:
Peter.Philippsen{at}unibas.ch.
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