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

S. cerevisiae cells acidify when they experience stressful temperatures. In addition, newly-translated proteins are thought to misfold, triggering the heat shock response. To determine whether heat-shock associated acidification and translation state are important for the cellular response, we manipulated intracellular pH, blocked translation, 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) or mock treated (RT for 20 minutes) in media where acidification was allowed or blocked, with translation ongoing or blocked; samples were collected just after treatment.