Detection of epigenetic mutagens including anthracene-derived compounds using yeast FLO1 promoter GFP reporter gene assay

Abstract

Recently, we have reported that the FLO1-mediated flocculation levels of yeast are affected by an epigenetic mutagen, alizarin. Alizarin promoted flocculation and reduced the bulk levels of histone H3 in yeast cells. Since alizarin has been known to possess carcinogenesis-promoting properties, it is important to estimate the effect of alizarin-related compounds on epigenome as measured by the flocculation of yeast. In this study, we examined the effects of two anthracene-derived compounds other than alizarin on the flocculation level of yeast. Purpurin significantly promoted the flocculation in a dose-dependent manner. While, quinizarin had a weaker promoting effect than purpurin. The strain treated with purprin showed FLO1 mRNA upregulation and reduced histone H3 expression similarly to alizarin. We also confirmed that the purprin-treated cells frequently exhibited abnormally shaped nuclei. Moreover, fluorescence intensities of green fluorescent protein (GFP) reporter under the FLO1 promoter control were dose-dependently increased by purprin and alizarin in the yeast. Taken together, these results suggest that the GFP reporter gene system utilising the FLO1 promoter is useful for the detection of epigenetic mutagens including anthracene-derived compounds.