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 Klobu
níková,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]