Figure 2.

msn5 mutants activate Crz1p-dependent transcription in a cnb1Δ background and alter Crz1p subcellular localization. (A) Strains harboring an integrated CDRE::lacZ reporter were grown to log phase at 30°C and harvested. β-Galactosidase activity was measured for cnb1Δ (DD12), msn5-11cnb1Δ (ASY11), and msn5Δ cnb1Δ (LBY196) strains. For each strain two cell extracts were prepared and assayed in triplicate. The standard deviation is the error between the samples. (B) WT (ASY472), cnb1Δ (ASY475), and msn5Δ cnb1Δ (LBY172) cells expressing GFP–Crz1p (AMS463) were grown to log phase at 21°C. Cells were treated with 200 mM CaCl₂, and GFP–Crz1p was visualized using fluorescence microscopy. Bar, 20 μm. (C) Msn5p does not affect Crz1p phosphorylation state. Whole cell extracts from wild-type (YPH499), cnb1Δ (DD12), and msn5Δ (ASY788) strains containing HA–Crz1p (AMS446) were treated with or without 2 μg/mL FK520 and analyzed by Western blot using anti-HA antibody.