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MBC in Press, published online ahead of print June 20, 2002
Mol. Biol. Cell 10.1091/mbc.E02-01-0003

A more recent version of this article appeared on August 1, 2002
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Submitted on January 7, 2002
Revised on May 20, 2002
Accepted on May 31, 2002

ß-Tubulin C354 Mutations that Severely Decrease Microtubule Dynamics do not Prevent Nuclear Migration in Yeast

Mohan L. Gupta Jr.1, Claudia J. Bode1, Douglas A. Thrower2, Chad G. Pearson2, Kathy A. Suprenant1, Kerry S. Bloom2, and Richard H. Himes1*

1 Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas, 66045
2 Department of Biology, University of North Carolina, Chapel Hill, North Carolina, 27599

* Corresponding author. E-mail address: himes{at}ku.edu.

Microtubule dynamics are influenced by interactions of microtubules with cellular factors and by changes in the primary sequence of the tubulin molecule. Mutations of yeast ß-tubulin C354, which is located near the binding site of some antimitotic compounds, reduce microtubule dynamicity greater than 90% in vivo and in vitro. The resulting intrinsically stable microtubules allowed us to determine which, if any, cellular processes are dependent on dynamic microtubules. The average number of cytoplasmic microtubules decreased from 3 in wild-type to 1 in mutant cells. The single microtubule effectively located the bud site before bud emergence. Although spindles were positioned near the bud neck at the onset of anaphase, the mutant cells were deficient in pre-anaphase spindle alignment along the mother-bud axis. Spindle microtubule dynamics and spindle elongation rates were also severely depressed in the mutants. The pattern and extent of cytoplasmic microtubule dynamics modulation through the cell cycle may reveal the minimum dynamic properties required to support growth. The ability to alter intrinsic microtubule dynamics and determine the in vivo phenotype of cells expressing the mutant tubulin provides a critical advance in assessing the dynamic requirements of an essential gene function.




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