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MBC in Press, published online ahead of print February 22, 2002
Mol. Biol. Cell 10.1091/mbc.01-07-0338

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Submitted on July 2, 2001
Revised on December 10, 2001
Accepted on January 14, 2002

Reorientation of mispositioned spindles in the short astral microtubule mutant spc72{Delta} is dependent on the spindle pole body outer plaque and the Kar3 motor protein

Dominic Hoepfner1, Florian Schaerer1, Arndt Brachat2, Achim Wach3, and Peter Philippsen1*

1 Lehrstuhl für Angewandte Mikrobiologie, Biozentrum, Universität Basel, Klingelbergstr.70, CH-4056 Basel, Switzerland
2 Lehrstuhl für Angewandte Mikrobiologie, Biozentrum, Universität Basel, Klingelbergstr.70, CH-4056 Basel, Switzerland (present address: Novartis Oncology, CH-4002 Basel, Switzerland)
3 Lehrstuhl für Angewandte Mikrobiologie, Biozentrum, Universität Basel, Klingelbergstr.70, CH-4056 Basel, Switzerland (present address: Bureco Corp., CH-4123 Allschwil, Switzerland)

* Corresponding author. E-mail address: peter.philippsen{at}unibas.ch.

Nuclear migration and positioning in Saccharomyces cerevisiae depends on long astral microtubules emanating from the spindle pole bodies (SPB). Here we show by in-vivo fluorescence microscopy that cells lacking Spc72, the SPB receptor of the cytoplasmic {gamma}-tubulin complex, can only generate very short (less than 1 µm) and unstable astral microtubules. Consequently, nuclear migration to the bud neck and orientation of the anaphase spindle along the mother-bud axis are absent in these cells. However, SPC72 deletion is not lethal because elongated but misaligned spindles can frequently reorient in mother cells permitting delayed but otherwise correct nuclear segregation. High-resolution time-lapse sequences revealed that this spindle reorientation was most likely accomplished by cortex interactions of the very short astral microtubules. In addition, a set of double mutants suggested that reorientation was dependent on the SPB outer plaque and the astral microtubule motor function of Kar3 but not Kip2/Kip3/Dhc1, or the cortex components Kar9/Num1. Our observations suggest that Spc72 is required for astral microtubule formation at the SPB half-bridge and for stabilization of astral microtubules at the SPB outer plaque. In addition, our data excludes involvement of Spc72 in spindle formation and elongation functions.




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A. S. Gladfelter, A. K. Hungerbuehler, and P. Philippsen
Asynchronous nuclear division cycles in multinucleated cells
J. Cell Biol., January 30, 2006; 172(3): 347 - 362.
[Abstract] [Full Text] [PDF]


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