FIG. 5.
The phenotypes of avo3ts mutants can be partially rescued by deletion of specific RhoGAP-encoding genes. (A) Deletion of SAC7 partially rescues the temperature sensitivity of avo3ts mutants. Tenfold serial dilutions of different yeast cultures were spotted onto YPD plates and incubated at different temperatures until colonies appeared. (B) The cell wall integrity of yeast strains carrying different GAP gene deletions was assessed by the trypan blue assay as described in Materials and Methods. The percentage of trypan blue-stained cells represents the extent of the cell wall defect of each strain. Shown are means ± standard deviations of results from three independent experiments. Statistical data were derived from Student's t test; each deletion strain was compared to its corresponding strain without any GAP gene deletion (No del). ***, P < 0.001; **, P < 0.005; *, P < 0.05. (C) Actin distribution in yeast strains carrying different GAP gene deletions was examined by TRITC-phalloidin staining and fluorescence microscopy. Small budded cells with four or more actin patches in the mother cell were counted as cells with abnormal actin distribution. Shown are means ± standard deviations of results from three independent experiments. Statistical data were obtained as for panel B.