![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 9, Issue 2, 523-543, February 1998
and
*Department of Molecular Genetics, University and
Biocenter of Vienna, A-1030 Vienna, Austria;
Overexpression of the yeast Pdr5 ATP-binding cassette transporter
leads to pleiotropic drug resistance to a variety of structurally unrelated cytotoxic compounds. To identify Pdr5 residues involved in
substrate recognition and/or drug transport, we used a combination of
random in vitro mutagenesis and phenotypic screening to isolate novel
mutant Pdr5 transporters with altered substrate specificity. A plasmid
library containing randomly mutagenized PDR5 genes was transformed into appropriate drug-sensitive yeast cells followed by
phenotypic selection of Pdr5 mutants. Selected mutant Pdr5 transporters
were analyzed with respect to their expression levels, subcellular
localization, drug resistance profiles to cycloheximide, rhodamines, antifungal azoles, steroids, and sensitivity to the inhibitor FK506. DNA sequencing of six PDR5 mutant genes
identified amino acids important for substrate recognition, drug
transport, and specific inhibition of the Pdr5 transporter. Mutations
were found in each nucleotide-binding domain, the transmembrane domain 10, and, most surprisingly, even in predicted extracellular hydrophilic loops. At least some point mutations identified appear to influence folding of Pdr5, suggesting that the folded structure is a major substrate specificity determinant. Surprisingly, a S1360F exchange in
transmembrane domain 10 not only caused limited substrate specificity, but also abolished Pdr5 susceptibility to inhibition by the
immunosuppressant FK506. This is the first report of a mutation in a
yeast ATP-binding cassette transporter that allows for the functional
separation of substrate transport and inhibitor susceptibility.
Department
of Biochemistry, Biozentrum, University of Basel, CH-4506 Basel,
Switzerland; and
Institut de Microbiologie, Centre
Hospitalier Universitaire Vaudois, CH-1011 Lausanne, Switzerland
This article has been cited by other articles:
|
|
 |
|
  D. Dietrich, H. Schmuths, C. D. M. Lousa, J. M. Baldwin, S. A. Baldwin, A. Baker, F. L. Theodoulou, and M. J. Holdsworth Mutations in the Arabidopsis Peroxisomal ABC Transporter COMATOSE Allow Differentiation between Multiple Functions In Planta: Insights from an Allelic Series Mol. Biol. Cell, January 1, 2009; 20(1): 530 - 543. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Ernst, P. Kueppers, C. M. Klein, T. Schwarzmueller, K. Kuchler, and L. Schmitt A mutation of the H-loop selectively affects rhodamine transport by the yeast multidrug ABC transporter Pdr5 PNAS, April 1, 2008; 105(13): 5069 - 5074. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. B. Mason, K. E. Allen, and C. W. Slayman Effects of C-terminal Truncations on Trafficking of the Yeast Plasma Membrane H+-ATPase J. Biol. Chem., August 18, 2006; 281(33): 23887 - 23898. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A.-K. Souid, C. Gao, L. Wang, E. Milgrom, and W.-C. W. Shen ELM1 Is Required for Multidrug Resistance in Saccharomyces cerevisiae Genetics, August 1, 2006; 173(4): 1919 - 1937. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Saini, T. Prasad, N. A. Gaur, S. Shukla, S. Jha, S. S. Komath, L. A. Khan, Q. Mohd. R. Haq, and R. Prasad Alanine scanning of transmembrane helix 11 of Cdr1p ABC antifungal efflux pump of Candida albicans: identification of amino acid residues critical for drug efflux J. Antimicrob. Chemother., July 1, 2005; 56(1): 77 - 86. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Gnann, J. R. Riordan, and D. H. Wolf Cystic Fibrosis Transmembrane Conductance Regulator Degradation Depends on the Lectins Htm1p/EDEM and the Cdc48 Protein Complex in Yeast Mol. Biol. Cell, September 1, 2004; 15(9): 4125 - 4135. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Shukla, S. V. Ambudkar, and R. Prasad Substitution of threonine-1351 in the multidrug transporter Cdr1p of Candida albicans results in hypersusceptibility to antifungal agents and threonine-1351 is essential for synergic effects of calcineurin inhibitor FK520 J. Antimicrob. Chemother., July 1, 2004; 54(1): 38 - 45. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Niimi, D. R. K. Harding, R. Parshot, A. King, D. J. Lun, A. Decottignies, M. Niimi, S. Lin, R. D. Cannon, A. Goffeau, et al. Chemosensitization of Fluconazole Resistance in Saccharomyces cerevisiae and Pathogenic Fungi by a D-Octapeptide Derivative Antimicrob. Agents Chemother., April 1, 2004; 48(4): 1256 - 1271. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. J. Rodriguez-Hernandez, I. Sanchez-Perez, R. Gil-Mascarell, A. Rodriguez-Afonso, A. Torres, R. Perona, and J. R. Murguia The Immunosuppressant FK506 Uncovers a Positive Regulatory Cross-talk between the Hog1p and Gcn2p Pathways J. Biol. Chem., September 5, 2003; 278(36): 33887 - 33895. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. K. Mukherjee, J. Chandra, D. M. Kuhn, and M. A. Ghannoum Mechanism of Fluconazole Resistance in Candida albicans Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols Infect. Immun., August 1, 2003; 71(8): 4333 - 4340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Gauthier, S. Weber, A.-M. Alarco, O. Alqawi, R. Daoud, E. Georges, and M. Raymond Functional Similarities and Differences between Candida albicans Cdr1p and Cdr2p Transporters Antimicrob. Agents Chemother., May 1, 2003; 47(5): 1543 - 1554. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Golin, S. V. Ambudkar, M. M. Gottesman, A. D. Habib, J. Sczepanski, W. Ziccardi, and L. May Studies with Novel Pdr5p Substrates Demonstrate a Strong Size Dependence for Xenobiotic Efflux J. Biol. Chem., February 14, 2003; 278(8): 5963 - 5969. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Mukhopadhyay, A. Kohli, and R. Prasad Drug Susceptibilities of Yeast Cells Are Affected by Membrane Lipid Composition Antimicrob. Agents Chemother., December 1, 2002; 46(12): 3695 - 3705. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Staleva, P. Manga, and S. J. Orlow Pink-eyed Dilution Protein Modulates Arsenic Sensitivity and Intracellular Glutathione Metabolism Mol. Biol. Cell, December 1, 2002; 13(12): 4206 - 4220. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-i. Wada, M. Niimi, K. Niimi, A. R. Holmes, B. C. Monk, R. D. Cannon, and Y. Uehara Candida glabrata ATP-binding Cassette Transporters Cdr1p and Pdh1p Expressed in a Saccharomyces cerevisiae Strain Deficient in Membrane Transporters Show Phosphorylation-dependent Pumping Properties J. Biol. Chem., November 22, 2002; 277(48): 46809 - 46821. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Falcon-Perez, M. Martinez-Burgos, J. Molano, M. J. Mazon, and P. Eraso Domain Interactions in the Yeast ATP Binding Cassette Transporter Ycf1p: Intragenic Suppressor Analysis of Mutations in the Nucleotide Binding Domains J. Bacteriol., August 15, 2001; 183(16): 4761 - 4770. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. J. Chen Activity of the Kluyveromyces lactis Pdr5 Multidrug Transporter Is Modulated by the Sit4 Protein Phosphatase J. Bacteriol., July 1, 2001; 183(13): 3939 - 3948. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Marchetti, J. M. Entenza, D. Sanglard, J. Bille, M. P. Glauser, and P. Moreillon Fluconazole plus Cyclosporine: a Fungicidal Combination Effective against Experimental Endocarditis Due to Candida albicans Antimicrob. Agents Chemother., November 1, 2000; 44(11): 2932 - 2938. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Sitcheran, R. Emter, A. Kralli, and K. R. Yamamoto A Genetic Analysis of Glucocorticoid Receptor Signaling: Identification and Characterization of Ligand-Effect Modulators in Saccharomyces cerevisiae Genetics, November 1, 2000; 156(3): 963 - 972. [Abstract] [Full Text]
|
|
|
|
|
|
O. Marchetti, P. Moreillon, M. P. Glauser, J. Bille, and D. Sanglard Potent Synergism of the Combination of Fluconazole and Cyclosporine in Candida albicans Antimicrob. Agents Chemother., September 1, 2000; 44(9): 2373 - 2381. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. J. Chen, B. E. Bauer, K. Kuchler, and G. D. Clark-Walker Positive and Negative Control of Multidrug Resistance by the Sit4 Protein Phosphatase in Kluyveromyces lactis J. Biol. Chem., May 12, 2000; 275(20): 14865 - 14872. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. E. Nau, L. R. Emerson, R. K. Martin, D. E. Kyle, D. F. Wirth, and M. Vahey Technical Assessment of the Affymetrix Yeast Expression GeneChip YE6100 Platform in a Heterologous Model of Genes That Confer Resistance to Antimalarial Drugs in Yeast J. Clin. Microbiol., May 1, 2000; 38(5): 1901 - 1908. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Del Poeta, M. C. Cruz, M. E. Cardenas, J. R. Perfect, and J. Heitman Synergistic Antifungal Activities of Bafilomycin A1, Fluconazole, and the Pneumocandin MK-0991/Caspofungin Acetate (L-743,873) with Calcineurin Inhibitors FK506 and L-685,818 against Cryptococcus neoformans Antimicrob. Agents Chemother., March 1, 2000; 44(3): 739 - 746. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. C. Freeman, S. J. Felts, D. O. Toft, and K. R. Yamamoto The p23 molecular chaperones act at a late step in intracellular receptor action to differentially affect ligand efficacies Genes & Dev., February 15, 2000; 14(4): 422 - 434. [Abstract] [Full Text]
|
|
|
|
|
|
J. Golin, A. Barkatt, S. Cronin, G. Eng, and L. May Chemical Specificity of the PDR5 Multidrug Resistance Gene Product of Saccharomyces cerevisiae Based on Studies with Tri-n-Alkyltin Chlorides Antimicrob. Agents Chemother., January 1, 2000; 44(1): 134 - 138. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Decottignies, G. Owsianik, and M. Ghislain Casein Kinase I-dependent Phosphorylation and Stability of the Yeast Multidrug Transporter Pdr5p J. Biol. Chem., December 24, 1999; 274(52): 37139 - 37146. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Sanglard, F. Ischer, D. Calabrese, P. A. Majcherczyk, and J. Bille The ATP Binding Cassette Transporter Gene CgCDR1 from Candida glabrata Is Involved in the Resistance of Clinical Isolates to Azole Antifungal Agents Antimicrob. Agents Chemother., November 1, 1999; 43(11): 2753 - 2765. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. E. Cardenas, M. C. Cruz, M. Del Poeta, N. Chung, J. R. Perfect, and J. Heitman Antifungal Activities of Antineoplastic Agents: Saccharomyces cerevisiae as a Model System To Study Drug Action Clin. Microbiol. Rev., October 1, 1999; 12(4): 583 - 611. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. L. Hogue, L. Kerby, and V. Ling A Mammalian Lysosomal Membrane Protein Confers Multidrug Resistance upon Expression in Saccharomyces cerevisiae J. Biol. Chem., April 30, 1999; 274(18): 12877 - 12882. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Plemper, J Bordallo, P. Deak, C Taxis, R Hitt, and D. Wolf Genetic interactions of Hrd3p and Der3p/Hrd1p with Sec61p suggest a retro-translocation complex mediating protein transport for ER degradation J. Cell Sci., January 11, 1999; 112(22): 4123 - 4134. [Abstract] [PDF]
|
|
|
|
|
|
R. K. Plemper, R. Egner, K. Kuchler, and D. H. Wolf Endoplasmic Reticulum Degradation of a Mutated ATP-binding Cassette Transporter Pdr5 Proceeds in a Concerted Action of Sec61 and the Proteasome J. Biol. Chem., December 4, 1998; 273(49): 32848 - 32856. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. W. Loo and D. M. Clarke Identification of Residues within the Drug-binding Domain of the Human Multidrug Resistance P-glycoprotein by Cysteine-scanning Mutagenesis and Reaction with Dibromobimane J. Biol. Chem., December 8, 2000; 275(50): 39272 - 39278. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. K. Hanson and J. W. Nichols Energy-dependent Flip of Fluorescence-labeled Phospholipids Is Regulated by Nutrient Starvation and Transcription Factors, PDR1 and PDR3 J. Biol. Chem., March 23, 2001; 276(13): 9861 - 9867. [Abstract] [Full Text] [PDF]
|
|
