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 Leber, R.
Right arrow Articles by Turnowsky, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Leber, R.
Right arrow Articles by Turnowsky, F.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, December 2003, p. 3890-3900, Vol. 47, No. 12
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.12.3890-3900.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Mechanism of Terbinafine Resistance in Saccharomyces cerevisiae

Regina Leber,1 Sandra Fuchsbichler,1 Vlasta Klobucníková,2 Natascha Schweighofer,1 Eva Pitters,1 Kathrin Wohlfarter,1 Mojca Lederer,1 Karina Landl,1 Christoph Ruckenstuhl,1 Ivan Hapala,2 and Friederike Turnowsky1*

Institute of Molecular Biology, Biochemistry and Microbiology, Karl-Franzens-Universität Graz, Graz, Austria,1 Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Ivanka pri Dunaji, Slovak Republic2

Received 10 April 2003/ Returned for modification 8 June 2003/ Accepted 20 August 2003

Ten mutants of the yeast Saccharomyces cerevisiae resistant to the antimycotic terbinafine were isolated after chemical or UV mutagenesis. Molecular analysis of these mutants revealed single base pair exchanges in the ERG1 gene coding for squalene epoxidase, the target of terbinafine. The mutants did not show cross-resistance to any of the substrates of various pleiotropic drug resistance efflux pumps tested. The ERG1 mRNA levels in the mutants did not differ from those in the wild-type parent strains. Terbinafine resistance was transmitted with the mutated alleles in gene replacement experiments, proving that single amino acid substitutions in the Erg1 protein were sufficient to confer the resistance phenotype. The amino acid changes caused by the point mutations were clustered in two regions of the Erg1 protein. Seven mutants carried the amino acid substitutions F402L (one mutant), F420L (one mutant), and P430S (five mutants) in the C-terminal part of the protein; and three mutants carried an L251F exchange in the central part of the protein. Interestingly, all exchanges identified involved amino acids which are conserved in the squalene epoxidases of yeasts and mammals. Two mutations that were generated by PCR mutagenesis of the ERG1 gene and that conferred terbinafine resistance mapped in the same regions of the Erg1 protein, with one resulting in an L251F exchange and the other resulting in an F433S exchange. The results strongly indicate that these regions are responsible for the interaction of yeast squalene epoxidase with terbinafine.

* Corresponding author. Mailing address: Institute of Molecular Biology, Biochemistry and Microbiology, Universitätsplatz 2, A-8010 Graz, Austria. Phone: 43 316 380 5623. Fax: 43 316 380 9898. E-mail: friederike.turnowsky{at}uni-graz.at.

Antimicrobial Agents and Chemotherapy, December 2003, p. 3890-3900, Vol. 47, No. 12
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.12.3890-3900.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ruckenstuhl, C., Poschenel, A., Possert, R., Baral, P. K., Gruber, K., Turnowsky, F. (2008). Structure-Function Correlations of Two Highly Conserved Motifs in Saccharomyces cerevisiae Squalene Epoxidase. Antimicrob. Agents Chemother. 52: 1496-1499 [Abstract] [Full Text]  
  • Meyer, V., Damveld, R. A., Arentshorst, M., Stahl, U., van den Hondel, C. A. M. J. J., Ram, A. F. J. (2007). Survival in the Presence of Antifungals: GENOME-WIDE EXPRESSION PROFILING OF ASPERGILLUS NIGER IN RESPONSE TO SUBLETHAL CONCENTRATIONS OF CASPOFUNGIN AND FENPROPIMORPH. J. Biol. Chem. 282: 32935-32948 [Abstract] [Full Text]  
  • Ruckenstuhl, C., Lang, S., Poschenel, A., Eidenberger, A., Baral, P. K., Kohut, P., Hapala, I., Gruber, K., Turnowsky, F. (2007). Characterization of Squalene Epoxidase of Saccharomyces cerevisiae by Applying Terbinafine-Sensitive Variants. Antimicrob. Agents Chemother. 51: 275-284 [Abstract] [Full Text]  
  • Rocha, E. M. F., Gardiner, R. E., Park, S., Martinez-Rossi, N. M., Perlin, D. S. (2006). A Phe389Leu Substitution in ErgA Confers Terbinafine Resistance in Aspergillus fumigatus.. Antimicrob. Agents Chemother. 50: 2533-2536 [Abstract] [Full Text]  
  • Osborne, C. S., Leitner, I., Hofbauer, B., Fielding, C. A., Favre, B., Ryder, N. S. (2006). Biological, Biochemical, and Molecular Characterization of a New Clinical Trichophyton rubrum Isolate Resistant to Terbinafine.. Antimicrob. Agents Chemother. 50: 2234-2236 [Abstract] [Full Text]  
  • Germann, M., Gallo, C., Donahue, T., Shirzadi, R., Stukey, J., Lang, S., Ruckenstuhl, C., Oliaro-Bosso, S., McDonough, V., Turnowsky, F., Balliano, G., Nickels, J. T. Jr. (2005). Characterizing Sterol Defect Suppressors Uncovers a Novel Transcriptional Signaling Pathway Regulating Zymosterol Biosynthesis. J. Biol. Chem. 280: 35904-35913 [Abstract] [Full Text]  
  • Osborne, C. S., Leitner, I., Favre, B., Ryder, N. S. (2005). Amino Acid Substitution in Trichophyton rubrum Squalene Epoxidase Associated with Resistance to Terbinafine. Antimicrob. Agents Chemother. 49: 2840-2844 [Abstract] [Full Text]  
  • Pasrija, R., Krishnamurthy, S., Prasad, T., Ernst, J. F., Prasad, R. (2005). Squalene epoxidase encoded by ERG1 affects morphogenesis and drug susceptibilities of Candida albicans. J Antimicrob Chemother 55: 905-913 [Abstract] [Full Text]  
  • Tsai, H.-F., Bard, M., Izumikawa, K., Krol, A. A., Sturm, A. M., Culbertson, N. T., Pierson, C. A., Bennett, J. E. (2004). Candida glabrata erg1 Mutant with Increased Sensitivity to Azoles and to Low Oxygen Tension. Antimicrob. Agents Chemother. 48: 2483-2489 [Abstract] [Full Text]  
  • Liu, W., May, G. S., Lionakis, M. S., Lewis, R. E., Kontoyiannis, D. P. (2004). Extra Copies of the Aspergillus fumigatus Squalene Epoxidase Gene Confer Resistance to Terbinafine: Genetic Approach to Studying Gene Dose-Dependent Resistance to Antifungals in A. fumigatus. Antimicrob. Agents Chemother. 48: 2490-2496 [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»