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Fig. 1. PtdIns(3)P accumulation is lethal in PI 3-phosphatase-deficient cells. (A) Deletion of the Vps34p PI 3-kinase complex component VPS30 rescues the lethal phenotype of ymr1 ${Delta}$ sjl2 ${Delta}$ sjl3 ${Delta}$ cells. ymr1 ${Delta}$ sjl2^ts sjl3 ${Delta}$ vps30 ${Delta}$ cells were transformed with either pRS415-VPS30 or an empty vector. Transformants were then streaked to 5-FOA plates lacking the amino acid leucine, and growth was scored after 5 days at 26°C. (B) Quantitative comparison of in vivo PtdIns(3)P levels in wild-type, ymr1 ${Delta}$ sjl2^ts sjl3 ${Delta}$ triple-mutant, ymr1 ${Delta}$ sjl2^ts sjl3 ${Delta}$ vps30 ${Delta}$ quadruple-mutant, and vps30 ${Delta}$ cells at 38°C. Levels of deacylated product corresponding to PtdIns(3)P are shown. Data represent the mean±s.d. of three independent experiments.

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