Copper potentiates azole antifungal activity in a way that does not involve complex formation.

To survive, fungal pathogens must acquire nutrient metals that are restricted by the host while also tolerating mechanisms of metal toxicity that are induced by the host. Given this dual vulnerability, we hypothesized that a pathogen's access to and control of essential yet potentially dangerous metal ions would affect fungal tolerance to antifungal drug stress. Here, we show that Candida albicans becomes sensitized to both Cu limitation and Cu elevation during exposure in liquid culture to the antifungal drug fluconazole, a widely prescribed antifungal agent. Spectroscopic data confirm that while fluconazole forms a complex with Cu(ii) in water, interactions of fluconazole with neither Cu(ii) nor Cu(i) are observed in the cell culture media used for the cellular assays. This result is further supported by growth assays in deletion strains that lack Cu import machinery. Overall, we establish that increases in Cu levels by as little as 40 nM over basal levels in the growth medium reduce tolerance of C. albicans to fluconazole in a way that does not require formation of a Cu-fluconazole complex. Rather, our data point to a more complex relationship between drug stress and Cu availability that gives rise to metal-mediated outcomes of drug treatment.