Measuring the dependence of the yeast heat shock response on intracellular pH during stress. Measuring the dependence of the yeast heat shock response on intracellular pH during stress

S. cerevisiae cells acidify when they experience stressful temperatures. To determine whether this heat-shock associated acidification is important for the cellular response, we manipulated intracellular pH, heat shocked cells, and sequenced the transcriptome. Overall design: Diploid yeast cells (BY4743 expressing 1 copy of SSA4 tagged genomically with mCherry and expressing pHluorin from the leu locus) were heat shocked (42C for 20 minutes) with and without ionophore treatment at 2 pH values; samples were collected just after treatment. Ionophore treatment at 6.8 mimicks normal cellular acidification; treatment at 7.4 prevents pH change during heat shock.