Figure 5

Synthetic rescue of the tor2-21^tsphenotype by myo1 Δ. Assay for viability of yeast strains by growth at 26°C and 37°C. Strains tested were wt (YJR24), myo1Δ, chs2 Δ, wt’ (JK9-3da), tor2 Δ ptor2^ts, wt ptor2^ts, myo1Δ ptor2^ts, chs2 Δ ptor2^ts, tor2 Δ pTOR2, myo1Δtor2 Δptor2^ts. A) Rescue of tor2–21^ts lethality at 37°C by myo1Δ in the YJR13 strain background (top) and SH121 strain background (bottom). B) Limiting dilution growth assay on agar medium measuring relative viability at 26°C and 37°C for tor2 Δ ptor2^ts, myo1Δtor2 Δptor2^ts, and myo1Δtor2 Δptor2^tspMYO1 strains. 10-fold dilutions are indicated at the top of the image (see Methods for details). C) Regulation of Slt2p phosphorylation in myo1Δ strains expressing the tor2–21^ts mutation at 37°C. Steady state levels of P-Slt2p in wt, myo1 Δ, tor2 Δ ptor2^ts, and myo1Δ ptor2^ts were analyzed by Western blot as described previously from cultures grown at 26°C and 37°C. Pgk1p was used as a loading control. Histograms show the ratio of the relative intensities of each P-Slt2p band and its Pgk1p loading control, averaged from duplicate experiments. Error bars represent STD Error Mean.