In vivo characterization of the drug resistance profile of the major ABC transporters and other components of the yeast pleiotropic drug resistance network.

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 PDRS, 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 Yor1p-independent 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 beta-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.

[1]  H. Jungwirth,et al.  Diazaborine Resistance in the Yeast Saccharomyces cerevisiae Reveals a Link between YAP1 and the Pleiotropic Drug Resistance Genes PDR1 andPDR3 * , 1997, The Journal of Biological Chemistry.

[2]  A. Goffeau,et al.  Molecular and phenotypic characterization of yeast PDR1 mutants that show hyperactive transcription of various ABC multidrug transporter genes , 1997, Molecular and General Genetics MGG.

[3]  A. Goffeau,et al.  Active efflux by multidrug transporters as one of the strategies to evade chemotherapy and novel practical implications of yeast pleiotropic drug resistance. , 1997, Pharmacology & therapeutics.

[4]  M. Raymond,et al.  AP1-mediated Multidrug Resistance in Saccharomyces cerevisiae Requires FLR1 Encoding a Transporter of the Major Facilitator Superfamily* , 1997, The Journal of Biological Chemistry.

[5]  K. Kuchler,et al.  Plasma Membrane Translocation of Fluorescent-labeled Phosphatidylethanolamine Is Controlled by Transcription Regulators, PDR1 and PDR3 , 1997, The Journal of cell biology.

[6]  I. Arias,et al.  A Yeast ATP-binding Cassette-type Protein Mediating ATP-dependent Bile Acid Transport* , 1997, The Journal of Biological Chemistry.

[7]  E. Balzi,et al.  Multidrug resistance in Aspergillus nidulans involves novel ATP-binding cassette transporters , 1997, Molecular and General Genetics MGG.

[8]  E. Bibi,et al.  MdfA, an Escherichia coli multidrug resistance protein with an extraordinarily broad spectrum of drug recognition , 1997, Journal of bacteriology.

[9]  V. Ling,et al.  Effect of quercetin on Hoechst 33342 transport by purified and reconstituted P-glycoprotein. , 1997, Biochemical pharmacology.

[10]  D. Sanglard,et al.  Cloning of Candida albicans genes conferring resistance to azole antifungal agents: characterization of CDR2, a new multidrug ABC transporter gene. , 1997, Microbiology.

[11]  André Goffeau,et al.  Complete inventory of the yeast ABC proteins , 1997, Nature Genetics.

[12]  A. F. Castro,et al.  Inhibition of drug transport by genistein in multidrug-resistant cells expressing P-glycoprotein. , 1997, Biochemical pharmacology.

[13]  D. Kelly,et al.  Resistance to fluconazole and cross‐resistance to amphotericin B in Candida albicans from AIDS patients caused by defective sterol Δ5,6‐desaturation , 1997, FEBS letters.

[14]  M H Saier,et al.  Multidrug‐Resistant Transport Proteins in Yeast: Complete Inventory and Phylogenetic Characterization of Yeast Open Reading Frames within the Major Facilitator Superfamily , 1997, Yeast.

[15]  J. Wingard,et al.  Isolation and characterization of fluconazole- and amphotericin B-resistant Candida albicans from blood of two patients with leukemia , 1997, Antimicrobial agents and chemotherapy.

[16]  M. Kołaczkowski,et al.  Anticancer Drugs, Ionophoric Peptides, and Steroids as Substrates of the Yeast Multidrug Transporter Pdr5p* , 1996, The Journal of Biological Chemistry.

[17]  R. Cannon,et al.  Multiple efflux mechanisms are involved in Candida albicans fluconazole resistance , 1996, Antimicrobial agents and chemotherapy.

[18]  K. Zhang,et al.  Inhibition of the efflux of glutathione S-conjugates by plant polyphenols. , 1996, Biochemical pharmacology.

[19]  G. Kruh,et al.  ATP-dependent transport of lipophilic cytotoxic drugs by membrane vesicles prepared from MRP-overexpressing HL60/ADR cells. , 1996, Biochemistry.

[20]  Piet Borst,et al.  MDR1 P-Glycoprotein Is a Lipid Translocase of Broad Specificity, While MDR3 P-Glycoprotein Specifically Translocates Phosphatidylcholine , 1996, Cell.

[21]  G. Kruh,et al.  Structure and in vitro substrate specificity of the murine multidrug resistance-associated protein. , 1996, Biochemistry.

[22]  A. Driessen,et al.  Multidrug resistance mediated by a bacterial homolog of the human multidrug transporter MDR1. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[23]  D. Sanglard,et al.  Susceptibilities of Candida albicans multidrug transporter mutants to various antifungal agents and other metabolic inhibitors , 1996, Antimicrobial agents and chemotherapy.

[24]  A. Driessen,et al.  Energetics and Mechanism of Drug Transport Mediated by the Lactococcal Multidrug Transporter LmrP* , 1996, The Journal of Biological Chemistry.

[25]  T. Hallstrom,et al.  Multiple Pdr1p/Pdr3p Binding Sites Are Essential for Normal Expression of the ATP Binding Cassette Transporter Protein-encoding Gene PDR5* , 1996, The Journal of Biological Chemistry.

[26]  A. Driessen,et al.  Multidrug resistance in Lactococcus lactis: evidence for ATP‐dependent drug extrusion from the inner leaflet of the cytoplasmic membrane. , 1996, The EMBO journal.

[27]  K. Tew,et al.  ATP-dependent uptake of natural product cytotoxic drugs by membrane vesicles establishes MRP as a broad specificity transporter. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[28]  W. Hillen,et al.  Tetracyclines: antibiotic action, uptake, and resistance mechanisms , 1996, Archives of Microbiology.

[29]  S. Kelly,et al.  Nonsterol related resistance in Ustilago maydis to the polyene antifungals, amphotericin B and nystatin. , 1996, Phytochemistry.

[30]  S. Kelly,et al.  Amphotericin B resistant isolates of Cryptococcus neoformans without alteration in sterol biosynthesis. , 1996, Journal of medical and veterinary mycology : bi-monthly publication of the International Society for Human and Animal Mycology.

[31]  K. Kuchler,et al.  The ATP‐binding cassette multidrug transporter Snq2 of Saccharomyces cerevisiae: a novel target for the transcription factors Pdr1 and Pdr3 , 1996, Molecular microbiology.

[32]  A. Cowman,et al.  The ATP-binding cassette (ABC) gene family of Plasmodium falciparum. , 1996, Parasitology today.

[33]  Richard G. W. Anderson,et al.  Acylation Targets Endothelial Nitric-oxide Synthase to Plasmalemmal Caveolae (*) , 1996, The Journal of Biological Chemistry.

[34]  F. Odds Resistance of clinically important yeasts to antifungal agents. , 1996, International journal of antimicrobial agents.

[35]  Jack Benner,et al.  Activation of Glycosylasparaginase , 1996, The Journal of Biological Chemistry.

[36]  M. Voet,et al.  Expression of an ATP-binding cassette transporter-encoding gene (YOR1) is required for oligomycin resistance in Saccharomyces cerevisiae , 1995, Molecular and cellular biology.

[37]  K. Kuchler,et al.  Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters , 1995, Antimicrobial agents and chemotherapy.

[38]  C. Jacq,et al.  Positive autoregulation of the yeast transcription factor Pdr3p, which is involved in control of drug resistance , 1995, Molecular and cellular biology.

[39]  A. Goffeau,et al.  Identification and Characterization of SNQ2, a New Multidrug ATP Binding Cassette Transporter of the Yeast Plasma Membrane (*) , 1995, The Journal of Biological Chemistry.

[40]  A. Willems,et al.  Studies on the transformation of intact yeast cells by the LiAc/SS‐DNA/PEG procedure , 1995, Yeast.

[41]  A. Goffeau,et al.  Yeast multidrug resistance: The PDR network , 1995, Journal of bioenergetics and biomembranes.

[42]  Fred Winston,et al.  Construction of a set of convenient saccharomyces cerevisiae strains that are isogenic to S288C , 1995, Yeast.

[43]  P. Rathod,et al.  Loss of function mutation in the yeast multiple drug resistance gene PDR5 causes a reduction in chloramphenicol efflux , 1994, Antimicrobial Agents and Chemotherapy.

[44]  J. Lankelma,et al.  Competitive inhibition by genistein and ATP dependence of daunorubicin transport in intact MRP overexpressing human small cell lung cancer cells. , 1994, Biochemical pharmacology.

[45]  A. Goffeau,et al.  Solubilization and characterization of the overexpressed PDR5 multidrug resistance nucleotide triphosphatase of yeast. , 1994, The Journal of biological chemistry.

[46]  H. Nikaido,et al.  Prevention of drug access to bacterial targets: permeability barriers and active efflux. , 1994, Science.

[47]  Jackson A. Como,et al.  Oral azole drugs as systemic antifungal therapy. , 1994, The New England journal of medicine.

[48]  H. M. Pinedo,et al.  Genistein modulates the decreased drug accumulation in non-P-glycoprotein mediated multidrug resistant tumour cells. , 1993, British Journal of Cancer.

[49]  I. Pastan,et al.  Fluorescent cellular indicators are extruded by the multidrug resistance protein. , 1993, The Journal of biological chemistry.

[50]  M. Kavallaris,et al.  Resistance to tetracycline, a hydrophilic antibiotic, is mediated by P-glycoprotein in human multidrug-resistant cells. , 1993, Biochemical and biophysical research communications.

[51]  H. Neu,et al.  The Crisis in Antibiotic Resistance , 1992, Science.

[52]  M. Montagu,et al.  The plasmid‐encoded chloramphenicol‐resistance protein of Rhodococcus fascians is homologous to the transmembrane tetracycline efflux proteins , 1992, Molecular microbiology.

[53]  B. E. Cohen,et al.  A sequential mechanism for the formation of aqueous channels by amphotericin B in liposomes. The effect of sterols and phospholipid composition. , 1992, Biochimica et biophysica acta.

[54]  R. Dudler,et al.  Structure of an mdr-like gene from Arabidopsis thaliana. Evolutionary implications. , 1992, The Journal of biological chemistry.

[55]  P. H. Roy,et al.  Characterization of the nonenzymatic chloramphenicol resistance (cmlA) gene of the In4 integron of Tn1696: similarity of the product to transmembrane transport proteins , 1991, Journal of bacteriology.

[56]  S. C. Falco,et al.  Cloning by gene amplification of two loci conferring multiple drug resistance in Saccharomyces. , 1990, Genetics.

[57]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[58]  A. Goffeau,et al.  The multidrug resistance gene PDR1 from Saccharomyces cerevisiae. , 1987, The Journal of biological chemistry.

[59]  Nancy Kleckner,et al.  A Method for Gene Disruption That Allows Repeated Use of URA3 Selection in the Construction of Multiply Disrupted Yeast Strains , 1987, Genetics.

[60]  S. Emr,et al.  The amino terminus of the yeast F1-ATPase beta-subunit precursor functions as a mitochondrial import signal , 1986, The Journal of cell biology.

[61]  Levchenko Ab,et al.  Yeast resistance to polyene antibiotics. II. An analysis of the sterol composition of Saccharomyces cerevisiae yeasts resistant to nystatin , 1984 .

[62]  G. H. Rank,et al.  Modification and inheritance of pleiotropic cross resistance and collateral sensitivity in Saccharomyces cerevisiae. , 1975, Genetics.

[63]  I. Pastan,et al.  Genetic analysis of the multidrug transporter. , 1995, Annual review of genetics.

[64]  M. Ouellette,et al.  New mechanisms of drug resistance in parasitic protozoa. , 1995, Annual review of microbiology.

[65]  I. Pastan,et al.  Biochemistry of multidrug resistance mediated by the multidrug transporter. , 1993, Annual review of biochemistry.

[66]  J. Holtum,et al.  Mechanisms and Agronomic Aspects of Herbicide Resistance , 1993 .

[67]  G. F. Ames,et al.  ATP-dependent transport systems in bacteria and humans: relevance to cystic fibrosis and multidrug resistance. , 1993, Annual review of microbiology.

[68]  L. W. Parks,et al.  Yeast sterols: yeast mutants as tools for the study of sterol metabolism. , 1985, Methods in enzymology.