Oxidative stress induces heat shock factor phosphorylation and HSF-dependent activation of yeast metallothionein gene transcription

Metallothioneins (MTs) are a class of low-molecular-weight, cysteine-rich metal-binding proteins that function in metal detoxification and oxidative stress protection. We demonstrate that transcription of the Saccharomyces cerevisiae MT gene CUP1 is strongly activated by the superoxide anion generator menadione. This activation is exacerbated in a strain lacking the gene encoding Cu, Zn superoxide dismutase (SOD1). CUP1 transcriptional activation by oxidative stress is dependent on a functional CUP1 promoter heat shock element (HSE) and the carboxy-terminal trans-activation domain of heat shock transcription factor (HSF). Furthermore, protection against oxidative stress conferred by CUP1 in a sod1 Delta strain requires HSF-mediated CUP1 transcription. Although in response to heat, HSE-mediated CUP1 transcription and HSF phosphorylation are transient, both CUP1 gene expression and HSF phosphorylation are sustained in response to oxidative stress. Moreover, the patterns of tryptic phosphopeptides resolved from HSF derived from cells subjected to heat shock or oxidative stress are distinct. These results demonstrate that transcription of the S. cerevisiae metallothionein gene under conditions of oxidative stress is mediated by HSF and that in response to distinct activation stimuli, HSF is differentially phosphorylated in a manner that parallels metallothionein gene transcription.