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Fig. 2. Characterization of a temperature-sensitive cdk5^ts mutant. (A) Growth of FB1 (wt) and cdk5^ts strains at 22°C and 34°C in solid complete medium (CMD). At low temperature the growth of control cells and cdk5^ts is similar. However, cdk5^ts cells show severe growth defects at 34°C. (B) Morphology of cdk5^ts cells after incubation at the restrictive temperature (34°C) for 8 hours. FITC-labeled wheat germ agglutinin (WGA) signal, which marks chitin-rich areas, is restricted to the growing tips in wild-type cells at any temperature, whereas in the cdk5^ts mutant the polar staining is lost at high temperature and WGA staining is concentrated in lateral walls. Nuclei were stained with DAPI. Bars, 10 µm. (C) After 12 hours at the restrictive temperature, cells carrying the cdk5^ts allele frequently lyse as evidenced by the presence of cell `ghosts' (arrows). By contrast, wild-type cells growing in the same conditions are cigar shaped and form polar buds (inset). Bars, 10 µm. (D) Sensitivity of cdk5^ts cells to cell wall inhibitors. Cultures of wild-type (FB1) and cdk5^ts strains were grown overnight at the permissive temperature, then diluted to an OD₆₀₀ of 1.0. Tenfold serial dilutions were made and 2 µl of each was plated in solid YPD medium treated with the indicated compounds. The plates were grown for 4 days at 22°C.

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