Figure 8.

Hog1 nuclear export and dephosphorylation depend on Hog1 kinase activity. Strain K4327 (hog1Δ) was transformed with centromer plasmids containing pVR65–WT (HOG1–GFP) or HOG1–GFP K/R (pVR65–K/R) and strain W303-1A (WT) with HOG1–GFP K/R (pVR65–K/R). After exposure to hyperosmotic conditions (selective media with 0.4 M NaCl, 5 min), cells were returned rapidly to iso-osmotic conditions (selective medium without NaCl), and localization of Hog1–GFP derivatives was determined after 5 min (A; positions of nuclei are indicated by arrows; bar, 5 μm), or samples were taken at the time points indicated, and protein extracts were prepared (B). Western blots were analyzed with anti-active p38 MAPK antibody. Asterisks indicate the positions of proteins that are nonspecifically recognized by anti-active human p38 antibody.