The determinants of heat‐shock element‐directed lacZ expression in Saccharomyces cerevisiae

Abstract

Heat-shock induction of heat-shock protein genes is due to a specific promoter element (the heat-shock element, HSE). This study used lacZ under HSE control (HSE-lacZ) to characterize HSE activity in Saccharomyces cerevisiae cells of different physiological states and differing genetic backgrounds. In batch fermentations HSE-lacZ induction by heat shock was maximal in exponential growth, and showed marked decline with the approach to stationary phase. Expression in the absence of heat shock was unaffected by growth phase, indicating that the growth-dependent expression of many yeast heat-shock genes uses promoter elements in addition to the HSE. Heat-induced expression was strongly influenced by the temperature at which cultures were grown. While basal, uninduced expression was constant during growth at different temperatures to 30 degrees C, induction by transfer to 39 degrees C was reduced by increases in growth temperature as low as 18-24 degrees C. Maximal HSE-lacZ induction (30- to 50-fold) was in cultures grown at low temperatures (18-24 degrees C), then heat shocked at 39 degrees C. Ethanol was a poor inducer. Mutations having little effect on HSE-lacZ expression included a respiratory petite; ubi4 (which inactivates the poly-ubiquitin gene); also ubc4 and ubc5 (which each inactivate one of the ubiquitin ligases involved in degradation of aberrant protein). pep4-3 increased both basal and induced beta-galactosidase about two-fold, probably because of slower turnover of this enzyme in pep4-3 strains.