This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Nakamura, K.
Right arrow Articles by Cannon, R. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nakamura, K.
Right arrow Articles by Cannon, R. D.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, December 2001, p. 3366-3374, Vol. 45, No. 12
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.12.3366-3374.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Functional Expression of Candida albicans Drug Efflux Pump Cdr1p in a Saccharomyces cerevisiae Strain Deficient in Membrane Transporters

Kenjirou Nakamura,1,2 Masakazu Niimi,1,3 Kyoko Niimi,1 Ann R. Holmes,1 Jenine E. Yates,1 Anabelle Decottignies,4 Brian C. Monk,1 Andre Goffeau,4 and Richard D. Cannon1,*

Department of Oral Sciences and Orthodontics, University of Otago, Dunedin, New Zealand1; General Research Institute, Nippon Dental University, Niigata,2 and Department of Bioactive Molecules, National Institute of Infectious Diseases, Tokyo,3 Japan; and Unité de Biochimie Physiologique, Université de Louvain, Louvain, Belgium4

Received 3 January 2001/Returned for modification 20 February 2001/Accepted 27 August 2001

Analysis of the transport functions of individual Candida albicans plasma membrane drug efflux pumps is hampered by the multitude of endogenous transporters. We have stably expressed C. albicans Cdr1p, the major pump implicated in multiple-drug-resistance phenotypes, from the genomic PDR5 locus in a Saccharomyces cerevisiae mutant (AD1-8u-) from which seven major transporters of the ATP-binding cassette (ABC) family have been deleted. High-level expression of Cdr1p, under the control of the S. cerevisiae PDR5 promoter and driven by S. cerevisiae Pdr1p transcriptional regulator mutation pdr1-3, was demonstrated by increased levels of mRNA transcription, increased levels of nucleoside triphosphatase activity, and immunodetection in plasma membrane fractions. S. cerevisiae AD1-8u- was hypersensitive to azole antifungals (the MICs at which 80% of cells were inhibited [MIC80s] were 0.625 µg/ml for fluconazole, <0.016 µg/ml for ketoconazole, and <0.016 µg/ml for itraconazole), whereas the strain (AD1002) that overexpressed C. albicans Cdr1p was resistant to azoles (MIC80s of fluconazole, ketoconazole, and itraconazole, 30, 0.5, and 4 µg/ml, respectively). Drug resistance correlated with energy-dependent drug efflux. AD1002 demonstrated resistance to a variety of structurally unrelated chemicals which are potential drug pump substrates. The controlled overexpression of C. albicans Cdr1p in an S. cerevisiae background deficient in other pumps allows the functional analysis of pumping specificity and mechanisms of a major ABC transporter involved in drug efflux from an important human pathogen.

* Corresponding author. Mailing address: Department of Oral Sciences and Orthodontics, University of Otago, P.O. Box 647, Dunedin, New Zealand. Phone: 64-3-479-7081. Fax: 64-3-479-0673. E-mail: richard.cannon{at}stonebow.otago.ac.nz.

Antimicrobial Agents and Chemotherapy, December 2001, p. 3366-3374, Vol. 45, No. 12
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.12.3366-3374.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Cannon, R. D., Lamping, E., Holmes, A. R., Niimi, K., Baret, P. V., Keniya, M. V., Tanabe, K., Niimi, M., Goffeau, A., Monk, B. C. (2009). Efflux-Mediated Antifungal Drug Resistance. Clin. Microbiol. Rev. 22: 291-321 [Abstract] [Full Text]  
  • Kolaczkowski, M., Kolaczkowska, A., Motohashi, N., Michalak, K. (2009). New High-Throughput Screening Assay To Reveal Similarities and Differences in Inhibitory Sensitivities of Multidrug ATP-Binding Cassette Transporters. Antimicrob. Agents Chemother. 53: 1516-1527 [Abstract] [Full Text]  
  • Sauna, Z. E., Bohn, S. S., Rutledge, R., Dougherty, M. P., Cronin, S., May, L., Xia, D., Ambudkar, S. V., Golin, J. (2008). Mutations Define Cross-talk between the N-terminal Nucleotide-binding Domain and Transmembrane Helix-2 of the Yeast Multidrug Transporter Pdr5: POSSIBLE CONSERVATION OF A SIGNALING INTERFACE FOR COUPLING ATP HYDROLYSIS TO DRUG TRANSPORT. J. Biol. Chem. 283: 35010-35022 [Abstract] [Full Text]  
  • Holmes, A. R., Lin, Y.-H., Niimi, K., Lamping, E., Keniya, M., Niimi, M., Tanabe, K., Monk, B. C., Cannon, R. D. (2008). ABC Transporter Cdr1p Contributes More than Cdr2p Does to Fluconazole Efflux in Fluconazole-Resistant Candida albicans Clinical Isolates. Antimicrob. Agents Chemother. 52: 3851-3862 [Abstract] [Full Text]  
  • Gulshan, K., Moye-Rowley, W. S. (2007). Multidrug Resistance in Fungi. Eukaryot Cell 6: 1933-1942 [Full Text]  
  • Cannon, R. D., Lamping, E., Holmes, A. R., Niimi, K., Tanabe, K., Niimi, M., Monk, B. C. (2007). Candida albicans drug resistance another way to cope with stress. Microbiology 153: 3211-3217 [Abstract] [Full Text]  
  • Lamping, E., Monk, B. C., Niimi, K., Holmes, A. R., Tsao, S., Tanabe, K., Niimi, M., Uehara, Y., Cannon, R. D. (2007). Characterization of Three Classes of Membrane Proteins Involved in Fungal Azole Resistance by Functional Hyperexpression in Saccharomyces cerevisiae. Eukaryot Cell 6: 1150-1165 [Abstract] [Full Text]  
  • Pasrija, R., Banerjee, D., Prasad, R. (2007). Structure and Function Analysis of CaMdr1p, a Major Facilitator Superfamily Antifungal Efflux Transporter Protein of Candida albicans: Identification of Amino Acid Residues Critical for Drug/H+ Transport. Eukaryot Cell 6: 443-453 [Abstract] [Full Text]  
  • Niimi, K., Maki, K., Ikeda, F., Holmes, A. R., Lamping, E., Niimi, M., Monk, B. C., Cannon, R. D. (2006). Overexpression of Candida albicans CDR1, CDR2, or MDR1 Does Not Produce Significant Changes in Echinocandin Susceptibility.. Antimicrob. Agents Chemother. 50: 1148-1155 [Abstract] [Full Text]  
  • Endres, C. J., Unadkat, J. D. (2005). Residues Met89 and Ser160 in the Human Equilibrative Nucleoside Transporter 1 Affect Its Affinity for Adenosine, Guanosine, S6-(4-Nitrobenzyl)-mercaptopurine Riboside, and Dipyridamole. Mol. Pharmacol. 67: 837-844 [Abstract] [Full Text]  
  • Monk, B. C., Niimi, K., Lin, S., Knight, A., Kardos, T. B., Cannon, R. D., Parshot, R., King, A., Lun, D., Harding, D. R. K. (2005). Surface-Active Fungicidal D-Peptide Inhibitors of the Plasma Membrane Proton Pump That Block Azole Resistance. Antimicrob. Agents Chemother. 49: 57-70 [Abstract] [Full Text]  
  • Niimi, M., Niimi, K., Takano, Y., Holmes, A. R., Fischer, F. J., Uehara, Y., Cannon, R. D. (2004). Regulated overexpression of CDR1 in Candida albicans confers multidrug resistance. J Antimicrob Chemother 54: 999-1006 [Abstract] [Full Text]  
  • Ton, V.-K., Rao, R. (2004). Functional expression of heterologous proteins in yeast: insights into Ca2+ signaling and Ca2+-transporting ATPases. Am. J. Physiol. Cell Physiol. 287: C580-C589 [Abstract] [Full Text]  
  • Niimi, K., Harding, D. R. K., Parshot, R., King, A., Lun, D. J., Decottignies, A., Niimi, M., Lin, S., Cannon, R. D., Goffeau, A., Monk, B. C. (2004). Chemosensitization of Fluconazole Resistance in Saccharomyces cerevisiae and Pathogenic Fungi by a D-Octapeptide Derivative. Antimicrob. Agents Chemother. 48: 1256-1271 [Abstract] [Full Text]  
  • Leber, R., Fuchsbichler, S., Klobucnikova, V., Schweighofer, N., Pitters, E., Wohlfarter, K., Lederer, M., Landl, K., Ruckenstuhl, C., Hapala, I., Turnowsky, F. (2003). Molecular Mechanism of Terbinafine Resistance in Saccharomyces cerevisiae. Antimicrob. Agents Chemother. 47: 3890-3900 [Abstract] [Full Text]  
  • Gauthier, C., Weber, S., Alarco, A.-M., Alqawi, O., Daoud, R., Georges, E., Raymond, M. (2003). Functional Similarities and Differences between Candida albicans Cdr1p and Cdr2p Transporters. Antimicrob. Agents Chemother. 47: 1543-1554 [Abstract] [Full Text]  
  • Mukhopadhyay, K., Kohli, A., Prasad, R. (2002). Drug Susceptibilities of Yeast Cells Are Affected by Membrane Lipid Composition. Antimicrob. Agents Chemother. 46: 3695-3705 [Abstract] [Full Text]  
  • Wada, S.-i., Niimi, M., Niimi, K., Holmes, A. R., Monk, B. C., Cannon, R. D., Uehara, Y. (2002). 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. 277: 46809-46821 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»