Different thresholds in the responses of two heat shock transcription factors, HSF1 and HSF3.

Avian cells express three HSF genes encoding a unique factor, HSF3, as well as homologues of mammalian HSF1 and HSF2. HSF1 is the major factor that mediates the heat shock signal in mammalian cells. We reported previously that cHSF3, as well as cHSF1, is activated by heat shock in chicken cells. In this study, we examined the functional differences between cHSF1 and cHSF3. Comparison of the heat-inducible DNA binding activity of cHSF1 with cHSF3 at various temperatures revealed that the latter was activated at higher temperatures than the former. At a mild heat shock, such as 41 degrees C, only cHSF1 was activated, whereas both cHSF1 and cHSF3 were activated following a severe heat shock at 45 degrees C. Heat-inducible nuclear translocation and trimerization were accompanied by DNA binding activity. We also observed that cHSF3 was activated by treating cells with higher concentrations of sodium arsenite compared to cHSF1. The DNA binding activity of cHSF3 by severe heat shock lasted for a longer period than that of cHSF1. Interestingly, the total amount of cHSF3 increased only upon severe heat shock, whereas that of HSF1 decreased. Substantial amounts of cHSF3 remained in the soluble fraction under severe heat shock, whereas cHSF1 rapidly moved to the insoluble fractions in that conditions. Comparison of transcriptional activity of the activation domains of cHSF1 and cHSF3 revealed that the activity of cHSF3 was as strong as that of cHSF1. These findings indicate that there are different thresholds for cHSF1 and cHSF3 and that cHSF3 is involved in the persistent and burst activation of stress genes upon severe stress in chicken cells. Pretreatment of cycloheximide elevated the threshold concentrations of arsenite of both factors. This suggests that denaturation of nascent polypeptides could be the first trigger for the activation of both factors, and the pathways for activation of cHSF1 and cHSF3 may be identical, or at least share some common mechanisms.