Table II. Class III vac mutants fail to maintain normal levels of PtdIns(3,5)P2 .
| PtdIns(3)P | PtdIns(3,5)P2 | |||
|---|---|---|---|---|
| + | − | + | − | |
| Wild-type | 68 ± 7 | 100 ± 10 | 71 ± 18 | 3.5 ± 1.1 |
| fab1-Δ1 | 126 ± 16 | 91 ± 3 | ≤1.5 | ≤1.5 |
| vac7-Δ1 | 145 ± 12 | 105 ± 2 | ≤1.5 | ≤1.5 |
| vac14-Δ1 | 120 ± 10 | 97 ± 12 | 1.7 ± 0.6 | ≤1.5 |
| fab1-Δ1 vac14-Δ1 | 141 ± 5 | 101 ± 4 | ≤1.5 | ≤1.5 |
Cells were grown in inositol-free media in the presence of 0.01 mCi/ml 3H inositol. 0.9 M NaCl was added to half the cultures for 10 min prior to extraction. Values were obtained as described in Table I and normalized to PtdIns(3)P no salt in wild-type. This peak was 2.0% of the total cpm eluted (80,000 cpm of 4 ± 106 total cpm). Values listed as ≤1.5 were areas where no peak above background was detected. The data (mean ± standard deviation) were calculated from results obtained in six independent experiments.