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Antimicrobial Agents and Chemotherapy, August 2005, p. 3101-3108, Vol. 49, No. 8
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.8.3101-3108.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Inactivation of the FCY2 Gene Encoding Purine-Cytosine Permease Promotes Cross-Resistance to Flucytosine and Fluconazole in Candida lusitaniae

Florence Chapeland-Leclerc, Julien Bouchoux, Abdelhak Goumar, Christiane Chastin, Jean Villard, and Thierry Noël*

Laboratoire des Sciences Végétales, UFR des Sciences Pharmaceutiques et Biologiques, Université Paris 5, 4 avenue de l'Observatoire, 75006 Paris, France

Received 27 January 2005/ Returned for modification 16 March 2005/ Accepted 21 April 2005

In a previous work, we described the possible relationship between a defect of purine-cytosine permease and the acquisition of a cross-resistance to the antifungal combination flucytosine (5FC) and fluconazole (FLC) in Candida lusitaniae (T. Noël, F. François, P. Paumard, C. Chastin, D. Brethes, and J. Villard, Antimicrob. Agents Chemother. 47:1275-1284, 2003). Using degenerate PCR and chromosome walking, we cloned two FCY2-like genes in C. lusitaniae. Northern blot analysis revealed that only one gene was expressed; it was named FCY2. The other one behaved as a pseudogene and was named FCY21. In order to better characterize the possible role of FCY2 in cross-resistance to 5FC-FLC, disruption experiments with auxotrophic strain 6936 ura3(D95V) FCY2 with an integrative vector carrying the URA3 gene and a partial sequence of the C. lusitaniae FCY2 gene were undertaken. Southern blot analysis revealed that homologous recombination events occurred in all transformants analyzed at rates of 50% at resident locus FCY2 and 50% at resident locus URA3, resulting in the genotypes ura3 fcy2::URA3 and ura3::URA3 FCY2, respectively. It was then demonstrated that only transformants harboring a disrupted fcy2 gene were resistant to 5FC, susceptible to FLC, and resistant to the 5FC-FLC combination. Finally, complementation experiments with a functional FCY2 gene restored 5FC and FLC susceptibilities to the wild-type levels. The results of this study provide molecular evidence that inactivation of the sole FCY2 gene promotes cross-resistance to the antifungal association 5FC-FLC in C. lusitaniae.

* Corresponding author. Mailing address: Laboratoire des Sciences Végétales, UFR des Sciences Pharmaceutiques et Biologiques, Université Paris 5, 4 avenue de l'Observatoire, 75006 Paris, France. Phone: (33) 1 53 73 96 41. Fax: (33) 1 53 73 96 40. E-mail: thierry.noel{at}univ-paris5.fr.

Antimicrobial Agents and Chemotherapy, August 2005, p. 3101-3108, Vol. 49, No. 8
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.8.3101-3108.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • McManus, B. A., Moran, G. P., Higgins, J. A., Sullivan, D. J., Coleman, D. C. (2009). A Ser29Leu Substitution in the Cytosine Deaminase Fca1p Is Responsible for Clade-Specific Flucytosine Resistance in Candida dubliniensis. Antimicrob. Agents Chemother. 53: 4678-4685 [Abstract] [Full Text]  
  • Florent, M., Noel, T., Ruprich-Robert, G., Da Silva, B., Fitton-Ouhabi, V., Chastin, C., Papon, N., Chapeland-Leclerc, F. (2009). Nonsense and Missense Mutations in FCY2 and FCY1 Genes Are Responsible for Flucytosine Resistance and Flucytosine-Fluconazole Cross-Resistance in Clinical Isolates of Candida lusitaniae. Antimicrob. Agents Chemother. 53: 2982-2990 [Abstract] [Full Text]  
  • Chapeland-Leclerc, F., Paccallet, P., Ruprich-Robert, G., Reboutier, D., Chastin, C., Papon, N. (2007). Differential Involvement of Histidine Kinase Receptors in Pseudohyphal Development, Stress Adaptation, and Drug Sensitivity of the Opportunistic Yeast Candida lusitaniae. Eukaryot Cell 6: 1782-1794 [Abstract] [Full Text]  
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