Figure 2.

Rescue of dna2–20 is TOR-specific. The dna2–20 mutant carrying pYDF88 (GAL1p→TOR1) was transformed with the following plasmids: pYDF18 (vector), pYDF66 (Tor1p), pJK3–3 (Tor2p) (Kunz et al., 1993), 324.34 (Vps34p) (Herman and Emr, 1990), YEp352-STT4 (Stt4p) (Yoshida et al., 1994), and YEp352-PIK1 (Pik1p) (Flanagan et al., 1993). All of these plasmids are 2μ-based and contain the genes under control of their endogenous promoters, except for the Tor1p expression construct that utilizes the GPD promoter (see MATERIALS AND METHODS). Transformants were selected on SCGal-leu minus the appropriate amino acid to also select for the various plasmids. Cells were scraped off plates (representing ∼100 colonies), resuspended in water, sonicated, and spotted onto YPD and YPGal plates. Ten thousand cells were spotted on the leftmost region with successive fivefold dilutions moving to the right. The plates were incubated for 3 d at 30°C and photographed.