The anti-fungal β-sitosterol targets the yeast oxysterol-binding protein Osh4

β-Sitosterol is a plant metabolite with a broad range of anti-fungal activity, however, this compound is not toxic against a few fungal species. The target of β-sitosterol and the nature of its selective toxicity are not yet clear. Using a yeast model system and taking advantage of molecular biology and computational approaches, we identify the target and explain why β-sitosterol is not toxic against some fungal pathogens.

β-Sitosterol (200 μg mL^-1 ) is toxic against yeast cells expressing only Osh4 (an oxysterol-binding protein) and harbouring a upc2-1 mutation (which enables sterol uptake), but not against yeast strains expressing all seven Osh proteins and harbouring a upc2-1 mutation. Furthermore, β-sitosterol is not toxic against yeast strains without the upc2-1 mutation irrespective of the number of Osh proteins being expressed. The deletion of COQ1 (a gene known to be highly induced upon deletion of OSH4) enhances the toxicity of β-sitosterol in yeast cells expressing only Osh4 and harbouring the upc2-1 mutation. Molecular modelling suggests that β-sitosterol binds to Osh4 and the binding mode is similar to the binding of cholesterol to Osh4.

Our results indicate that the concentrations of β-sitosterol, and Osh4, as well as its homologues within cells, are most likely the main determinants of β-sitosterol toxicity. Furthermore, some fungal species do not take up sterols, e.g. Saccharomyces cerevisiae, under aerobic conditions. Therefore, sterol uptake may also contribute to the β-sitosterol anti-fungal effect. These findings enable predicting the toxicity of β-sitosterol against plant fungal pathogens. © 2019 Society of Chemical Industry.