Synthetic rescue of the tor2-21ts phenotype bymyo1Δ. Assay for viability of yeast strains by growth at 26°C and 37°C. Strains tested were wt (YJR24), myo1Δ, chs2Δ, wt’ (JK9-3da), tor2Δ ptor2ts, wt ptor2ts, myo1Δ ptor2ts, chs2Δ ptor2ts, tor2Δ pTOR2, myo1Δtor2Δptor2ts. A) Rescue of tor2–21ts 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 tor2Δ ptor2ts, myo1Δtor2Δptor2ts, and myo1Δtor2Δptor2tspMYO1 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 tor2–21ts mutation at 37°C. Steady state levels of P-Slt2p in wt, myo1Δ, tor2Δ ptor2ts, and myo1Δ ptor2ts 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.
Pagán-Mercado et al. BMC Cell Biology 2012 13:13 doi:10.1186/1471-2121-13-13