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Vol. 9, Issue 7, 1757-1771, July 1998
-Tubulin and Nonessential 
2-Tubulin Genes as
Fission Yeast Polarity Mutants
Laboratory of Cell Regulation, Imperial Cancer Research Fund,
London WC2A 3PX, United Kingdom
We have screened for temperature-sensitive (ts) fission yeast
mutants with altered polarity (alp1-15). Genetic
analysis indicates that alp2 is allelic to
atb2 (one of two 
-tubulin genes) and alp12 to nda3 (the single 
-tubulin
gene). atb2+ is nonessential, and the ts
atb2 mutations we have isolated are dominant as
expected. We sequenced two alleles of ts atb2 and one
allele of ts nda3. In the ts atb2
mutants, the mutated residues (G246D and C356Y) are found at the
longitudinal interface between /-heterodimers, whereas in ts
nda3 the mutated residue (Y422H) is situated in the
domain located on the outer surface of the microtubule. The ts
nda3 mutant is highly sensitive to altered gene dosage
of atb2+; overexpression of
atb2+ lowers the restrictive temperature,
and, conversely, deletion rescues ts. Phenotypic analysis shows that
contrary to undergoing mitotic arrest with high viability via the
spindle assembly checkpoint as expected, ts nda3 mutants
execute cytokinesis and septation and lose viability. Therefore, it
appears that the ts nda3 mutant becomes temperature
lethal because of irreversible progression through the cell cycle in
the absence of activating the spindle assembly checkpoint pathway.
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