Anticancer Drugs, Ionophoric Peptides, and Steroids as Substrates of the Yeast Multidrug Transporter Pdr5p

Pdr5p is the yeast Saccharomyces cerevisiae ATP-binding cassette transporter conferring resistance to several unrelated drugs. Its high overproduction in Pdr1p transcription factor mutants allows us to study the molecular mechanism of multidrug transport and substrate specificity. We have developed...

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Bibliographic Details
Published in:The Journal of biological chemistry 1996-12, Vol.271 (49), p.31543-31548
Main Authors: Kolaczkowski, Marcin, Michel, van der Rest, Cybularz-Kolaczkowska, Anna, Soumillion, Jean-Philippe, Konings, Wil N., André, Goffeau
Format: Article
Language:English
Subjects:
Animals
Antineoplastic Agents - metabolism
ATP-Binding Cassette Transporters - metabolism
Daunorubicin - pharmacology
Desoxycorticosterone - pharmacology
Doxorubicin - pharmacology
Fungal Proteins - metabolism
Ionophores - metabolism
Kinetics
Membrane Proteins - metabolism
Phenotype
Progesterone - pharmacology
Rhodamine 123
Rhodamines - pharmacology
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
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Summary:Pdr5p is the yeast Saccharomyces cerevisiae ATP-binding cassette transporter conferring resistance to several unrelated drugs. Its high overproduction in Pdr1p transcription factor mutants allows us to study the molecular mechanism of multidrug transport and substrate specificity. We have developed new in vivo and in vitro assays of Pdr5p-mediated drug transport. We show that in spite of little sequence homology, and inverted topology in respect to that of mammalian P-glycoproteins, Pdr5p shares with them common substrates. Pdr5p extrudes rhodamines 6G and 123, from intact yeast cells in an energy-dependent manner. Plasma membrane preparations from a Pdr5p-overproducing strain exhibit ATP hydrolysis-dependent, osmotically sensitive rhodamine 6G fluorescence quenching. The quenching is competitively inhibited by micromolar concentrations of many anticancer drugs, such as vinblastine, vincristine, taxol, and verapamil, and of ionophoric peptides as well as steroids. In contrast, other anticancer drugs, like colchicine and some multidrug resistance modifiers, such as quinidine, exert noncompetitive inhibition. Our experimental system opens new possibilities for the analysis of structure-function relationship of multidrug transporter substrates and inhibitors.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.271.49.31543