FIGURE 1.

Constitutively active Ypk2 rescues phenotypes of avo3^ts mutants. Two avo3^ts mutants carrying the control vector (pRS424), or a plasmid expressing AVO3 (pTSS1) or YPK2^ΔN (pHC1) were examined in different assays. A, temperature sensitivity assay. 10-fold serial dilutions of cell suspensions were spotted on SC-Trp plates and incubated at the indicated temperatures until colonies formed. B, caffeine sensitivity assay. Cell suspensions were spotted on YPD plates or YPD plates containing 8 mm caffeine and incubated at 27 °C until colonies formed. C, trypan blue assy. Different cultures were diluted to A₆₀₀ = 0.15 and shifted to indicated temperatures for 6 h. The cells were stained with trypan blue and observed under the microscope. Shown are results from three independent experiments. **, p < 0.01; *, p < 0.05 (Student's t test). D, actin distribution assay. Log phase cultures were shifted to 37 °C for 3 h, fixed, and stained for F-actin using TRITC-phalloidin. Individual cells were examined by fluorescence and differential interference contrast microscopy. Small-budded cells showing four or more actin patches in the mother cells were scored as cells with aberrant actin distribution. Shown are the data from three independent experiments. **, p < 0.01; *, p < 0.05 (Student's t test). caff., caffeine; Vec, vector; aberr, aberrant.