In Vivo Characterization of the Drug Resistance Profile of the Major ABC Transporters and Other Components of the Yeast Pleiotropic Drug Resistance Network

ABSTRACT

Multidrug resistance (MDR) mediated by broad specificity transporters is one of the most important strategies used by pathogens, including cancer cells, to evade chemotherapy. In the yeast Saccharomyces cerevisiae, a complex pleiotropic drug resistance (PDR) network of genes involved in MDR is composed of the transcriptional regulators Pdr1p and Pdr3p, which activate expression of the ATP-binding cassette (ABC) MDR transporters-encoding genes PDR5, SNQ2, and YOR1 as well as other not yet identified genes. We have screened 349 toxic compounds in isogenic S. cerevisiae strains deleted of PDR5, SNQ2, or YOR1 in different combinations as well as both PDR1 and PDR3. The screen revealed extremely promiscuous, yet limited, and to a large extent overlapping but distinct drug resistance profiles of Pdr5p, Snq2p, and Yor1p. These ABC-MDR transporters mediated resistance to most currently available classes of clinically and agriculturally important fungicides and also to many antibiotics, herbicides, and others. Several classes of compounds were identified for the first time in the drug resistance spectrum of MDR transporters. These are fungicides, such as anilinopyrimidines, benzimidazoles, benzenedicarbonitriles, dithiocarbamates, guanidines, imidothiazoles, polyenes, pyrimidynyl carbinols, and strobilurine analogues; the urea derivative and anilide herbicides; flavonoids, several membrane lipids resembling detergents; and newly synthesized lysosomotropic aminoesters; as well as many others. Identification of compounds showing Pdr1p, Pdr3p-dependent, but Pdr5p-, Snq2p-, and Yor1pindependent toxicity, reflected in the case of rhodamine 6G, by efflux alterations, suggests the involvement of new drug resistance genes and is a first step toward their identification. The highly increased toxicity of bile acids toward the PDR1, PDR3 double disruptant together with the decreased level of BAT1 promoter dependent β-galactosidase activity suggest that the Bat1p ABC transporter is a new member of the PDR network. Our results may contribute to a better understanding of the mechanism of MDR, in particular in the pathogenic yeast Candida albicans. They also provide and indication of the physiological function of MDR transporters and suggest new approaches for the cloning of the mammalian bile acid transporters.