Negative interaction between TORC1 activity and the Wsc1p cell wall stress sensor. A) Western blot analysis of Npr1p electrophoretic mobility in wsc1Δ mutant strains (see methods for details). Whole cell protein extracts were prepared from myo1Δ(40 μg), wsc1Δ(20 μg) and myo1Δwsc1Δ(20 μg) strains expressing the HA-NPR1 as described. Reactivation of TORC1 is observed in a myo1Δwsc1Δ strain and dephosphorylation occurs upon Inhibition of TORC1 by rapamycin. A wsc1Δ strain shows down regulation of TORC1. B) Western blot analysis of P-Slt2p levels in wsc1Δ mutant strains. 50 μg whole cell protein extracts were analyzed per lane. 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 STDError mean. C) Limiting dilution growth assay on agar medium measuring relative viability of wt, myo1Δ, wsc1Δ and myo1Δwsc1Δ strains at 26°C. 10-fold dilutions are indicated at the top of the image (see Methods for details).
Pagán-Mercado et al. BMC Cell Biology 2012 13:13 doi:10.1186/1471-2121-13-13