Table 1.
svl mutant complementation groups
| Group | Class/allele | proCPY | proAPI | Ste3p | PC | PE |
|---|---|---|---|---|---|---|
| SVL | − | <5% | <5% | ≈25 min | 1.00 | 1.00 |
| svl2 | II/vps41 | ++++ | ++++ | >60 min | 0.81 | 0.49 |
| svl3 | I | <5% | <5% | ≈20 min | 1.60 | 0.97 |
| svl4 | I | ND | ND | ≈46 min | 1.01 | 1.35 |
| svl5 | III | ++ | ++++ | ≈24 min | 0.95 | 2.10 |
| svl6 | II/vps16 | ++++ | ++++ | >60 min | 2.36 | 2.53 |
| svl7 | I/fab1 | ND | ND | ≈28 min | 1.16 | 1.17 |
| svl8 | III | <5% | +++ | ≈58 min | 2.80* | 4.93* |
| svl9 | II | +++ | ++++ | ND | 2.69 | 1.25 |
| svl10 | I | ++++ | ++++ | >60 min | 1.17 | 1.53 |
| svl11 | I | +++ | ++++ | >60 min | 1.24 | 1.48 |
| svl12 | III | <5% | ++ | ≈15 min | 1.18 | 1.14 |
Mutant class I, large vacuoles; class II, fragmented vacuoles; and class III, punctate/diffuse structures. Alleles of previous mutants are indicated. For accumulation of proCPY and proAPI, each + sign represents 25% of the total from Western blot analysis. The turnover of Ste3p was performed as described in Materials and Methods and is reported in half times. Uptake of both phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was performed using BODIPY-labeled lipids and 10–30 cells were quantitated with digital image analysis. All values were expressed relative to SVL cells arbitrarily set to 1.00.
All svl mutants showed wild-type morphology for both phosphatidylcholine and phosphatidylethanolamine except svl8, which showed accumulation at the plasma membrane (see Fig. 3B).