Abstract

Tirapazamine (TPZ) is an anticancer drug that targets topoisomerase II. TPZ is preferentially active under hypoxic conditions. The drug itself is not harmful to cells; rather, it is reduced to a toxic radical species by an NADPH cytochrome P450 oxidoreductase. Under aerobic conditions, the toxic compound reacts with oxygen to revert back to TPZ and a much less toxic radical species. We have used yeast (Saccharomyces cerevisiae) as a model to better understand the mechanism of action of TPZ. Overexpression of NCP1, encoding the yeast ortholog of the human P450 oxidoreductase, results in greatly increased sensitivity to TPZ. Likewise, overexpression of TOP2 (encoding topoisomerase II) leads to hypersensitivity to TPZ, suggesting that topoisomerase II is also a target of TPZ in yeast. Thus, our data show that yeast mimics human cells in terms of TPZ sensitivity. We have performed robot-aided screens for altered sensitivity to TPZ using a collection of approximately 4600 haploid yeast deletion strains. We have identified 117 and 73 genes whose deletion results in increased or decreased resistance to TPZ, respectively. For example, cells lacking various DNA repair genes are hypersensitive to TPZ. In contrast, deletion of genes encoding some amino acid permeases results in cells that are resistant to TPZ. This suggests that permeases may be involved in intracellular uptake of TPZ. Our discoveries in yeast may lead to a better understanding of TPZ biology in humans.