Study gene expression specifically regulated by intrinsically active HOG1

To monitor changes in the transcriptome following activation of Hog1 per se, we used a previously established inducible expression system of intrinsically active Hog1 proteins the genes encoding HOG1WT, HOG1D170A and HOG1D170A+F318L, cloned downstream of the MET3 promoter, were introduced into hog1∆ cells . JBY13 cells harboring an ‘empty’ plasmid served as a control. The MET3 promoter is shut off when the growth media contains methionine, and is rapidly induced when methionine is removed.