Fig. 5

Indirect reporters of the level of PKA activity in the light-sensitive mutants. a Time-lapse micro-graphs of Msn2-GFP in wild type, hog1∆, and rps21B∆. b–e The total nuclear localization trace of each strain compared to the wild type. The gray area corresponds to the mean value of 15 individual experiments of the wild type strain plus/minus one standard deviation. Each wild type experiment contains 50–60 cells. The trace for each deletion strain is the combined response for > 99 cells (up to 170 cells) from 2 or 3 experiments on individual days. Data from the deletion strains have been smoothed using a spline smoothing algorithm in order to visualize the main trend for each individual strain. Msn2 nuclear localization for selected single deletion strains during continuous light exposure response has been divided into 4 groups. Many light-sensitive mutants display reduced Msn2-GFP nuclear localization in response to light. b Normal/wild type response. c Moderately reduced response where Msn2 spends less time in the nucleus. d Strongly reduced response, where Msn2p does not enter the nucleus and e early, but otherwise normal response, unique for pho5Δ. f Glycogen staining of cells from the indicated strains using iodine vapor. A representative result from at least two independent experiments is shown. In the top row, the wild-type transformed with a centromeric plasmid expressing hyperactive RAS2G19V (pRAS2V19) yielding constitutively high PKA activity (PKA ++), multicopy PDE2 (pPDE2; phosophodiesterase) resulting in low PKA activity (PKA −), or the corresponding vector controls (vector, intermediate PKA activity +) are shown. The three remaining rows display glycogen staining of the indicated light-sensitive deletion mutants classified as staining similar to the wild-type (wt glycogen), lower than the wild-type (low glycogen), or higher than the wild-type (high glycogen)