A screen of yeast respiratory mutants for sensitivity against the mycotoxin citrinin identifies the vacuolar ATPase as an essential factor for the toxicity mechanism

In countries with a hot climate the mycotoxin citrinin represents a serious problem in fungal food-poisoning. In humans the renal system is affected t he most and the mitochondrial respiratory chain was identified as a possibl e sensitive target for this toxin. In addition. citrinin has an antifungal activity that also inhibits the growth of the yeast Saccharomyces cerevisia e. So far the precise mode of action and the subcellular targets for citrin in have not been identified. Therefore, we decided to use the model organis m yeast for a genetic approach to identify genes that play a role in the se nsitivity against this mycotoxin. A large collection of conditional respira tory deficient yeast mutants was screened for sensitivity against citrinin. One special pel-ts mutant was identified that exhibited a higher sensitivi ty against citrinin. The genetic system of yeast allowed the isolation of t he respective wild-type gene. This yeast gene encodes the Vph2p subunit tha t is essential for the correct assembly of the vacuolar ATPase. Isolation o f the mutated gene and gene-disruption experiments of VPH2 and the partiall y overlapping small YKL118W gene verified this finding. The wild-type VPHL2 gene restores all defects of the mutants. In contrast to this, YKL118W gav e no complementation and the null mutant showed no phenotype. Thereby the y east vacuolar,ATPase was found to be important for the toxic effect of citr inin in yeast cells. The consequences of this finding for the molecular mec hanism of citrinin action and its relation to the mitochondrial respiratory chain are discussed.