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Vol. 12, Issue 7, 2119-2136, July 2001
-Tubulin Yields Diverse and Novel Phenotypes
*Department of Molecular Genetics, The Ohio State University,
Columbus, Ohio 43210; and §Howard Hughes Medical Institute
Molecular and Cell Biology Department and Life Science Division,
Lawrence Berkeley National Laboratory, University of California at
Berkeley, Berkeley, California 94720
We have created 41 clustered charged-to-alanine scanning mutations
of the mipA, 
-tubulin, gene of Aspergillus
nidulans and have created strains carrying these mutations by
two-step gene replacement and by a new procedure, heterokaryon gene
replacement. Most mutant alleles confer a wild-type phenotype, but
others are lethal or conditionally lethal. The conditionally lethal
alleles exhibit a variety of phenotypes under restrictive conditions. Most have robust but highly abnormal mitotic spindles and some have
abnormal cytoplasmic microtubule arrays. Two alleles appear to have
reduced amounts of -tubulin at the spindle pole bodies and
nucleation of spindle microtubule assembly may be partially inhibited.
One allele inhibits germ tube formation. The cold sensitivity of two
alleles is strongly suppressed by the antimicrotubule agents benomyl
and nocodazole and a third allele is essentially dependent on these
compounds for growth. Together our data indicate that -tubulin
probably carries out functions essential to mitosis and organization of
cytoplasmic microtubules in addition to its well-documented role in
microtubule nucleation. We have also placed our mutations on a model of
the structure of -tubulin and these data give a good initial
indication of the functionally important regions of the molecule.
Present addresses: NCI-Frederick Cancer
Research and Development Center, Bldg. 469/Room 140, Frederick, MD
21702;
The Scripps Research Institute, Department of
Cell Biology, MB-39, Room MBB201, 10550 North Torrey Pines Rd., La
Jolla, CA 92037.
Corresponding author. E-mail address:
oakley.2{at}osu.edu.
Online version of this article contains video material
for some figures. Online version is available at www.molbiolcell.org.
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