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 Mann, P. A.
Right arrow Articles by McNicholas, P. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mann, P. A.
Right arrow Articles by McNicholas, P. M.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, February 2003, p. 577-581, Vol. 47, No. 2
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.2.577-581.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Mutations in Aspergillus fumigatus Resulting in Reduced Susceptibility to Posaconazole Appear To Be Restricted to a Single Amino Acid in the Cytochrome P450 14{alpha}-Demethylase

Paul A. Mann, Raulo M. Parmegiani, Shui-Qing Wei, Cara A. Mendrick, Xin Li, David Loebenberg, Beth DiDomenico, Roberta S. Hare, Scott S. Walker, and Paul M. McNicholas*

Schering-Plough Research Institute, Kenilworth, New Jersey 07033

Received 25 September 2002/ Returned for modification 31 October 2002/ Accepted 19 November 2002

To better understand the molecular basis of posaconazole (POS) resistance in Aspergillus fumigatus, resistant laboratory isolates were selected. Spontaneous mutants arose at a frequency of 1 in 108 and fell into two susceptibility groups, moderately resistant and highly resistant. Azole resistance in A. fumigatus was previously associated with decreased drug accumulation. We therefore analyzed the mutants for changes in levels of transcripts of genes encoding efflux pumps (mdr1 and mdr2) and/or alterations in accumulation of [14C]POS. No changes in either pump expression or drug accumulation were detected. Similarly, there was no change in expression of cyp51A or cyp51B, which encode the presumed target site for POS, cytochrome P450 14{alpha}-demethylase. DNA sequencing revealed that each resistant isolate carried a single point mutation in residue 54 of cyp51A. Mutations at the same locus were identified in three clinical A. fumigatus isolates exhibiting reduced POS susceptibility but not in susceptible clinical strains. To verify that these mutations were responsible for the resistance phenotype, we introduced them into the chromosome of a POS-susceptible A. fumigatus strain under the control of the glyceraldehyde phosphate dehydrogenase promoter. The transformants exhibited reductions in susceptibility to POS comparable to those exhibited by the original mutants, confirming that point mutations in the cyp51A gene in A. fumigatus can confer reduced susceptibility to POS.

* Corresponding author. Mailing address: Schering-Plough Research Institute, 2015 Galloping Hill Rd., 4700, Kenilworth, NJ 07033. Phone: (908) 740-7644. Fax: (908) 740-3918. E-mail: paul.mcnicholas{at}spcorp.com.

Antimicrobial Agents and Chemotherapy, February 2003, p. 577-581, Vol. 47, No. 2
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.2.577-581.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Araujo, R., Espinel-Ingroff, A. (2009). Comparison of Assessment of Oxygen Consumption, Etest, and CLSI M38-A2 Broth Microdilution Methods for Evaluation of the Susceptibility of Aspergillus fumigatus to Posaconazole. Antimicrob. Agents Chemother. 53: 4921-4923 [Abstract] [Full Text]  
  • Pfaller, M. A., Diekema, D. J., Ghannoum, M. A., Rex, J. H., Alexander, B. D., Andes, D., Brown, S. D., Chaturvedi, V., Espinel-Ingroff, A., Fowler, C. L., Johnson, E. M., Knapp, C. C., Motyl, M. R., Ostrosky-Zeichner, L., Sheehan, D. J., Walsh, T. J., for the Clinical and Laboratory Standards Institut, (2009). Wild-Type MIC Distribution and Epidemiological Cutoff Values for Aspergillus fumigatus and Three Triazoles as Determined by the Clinical and Laboratory Standards Institute Broth Microdilution Methods. J. Clin. Microbiol. 47: 3142-3146 [Abstract] [Full Text]  
  • 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]  
  • Morris, M. I. (2009). Posaconazole: A new oral antifungal agent with an expanded spectrum of activity. Am J Health Syst Pharm 66: 225-236 [Abstract] [Full Text]  
  • Arendrup, M. C., Perkhofer, S., Howard, S. J., Garcia-Effron, G., Vishukumar, A., Perlin, D., Lass-Florl, C. (2008). Establishing In Vitro-In Vivo Correlations for Aspergillus fumigatus: the Challenge of Azoles versus Echinocandins. Antimicrob. Agents Chemother. 52: 3504-3511 [Abstract] [Full Text]  
  • Rachwalski, E. J, Wieczorkiewicz, J. T, Scheetz, M. H (2008). Posaconazole: An Oral Triazole with an Extended Spectrum of Activity. The Annals of Pharmacotherapy 42: 1429-1438 [Abstract] [Full Text]  
  • Pfaller, M. A., Messer, S. A., Boyken, L., Rice, C., Tendolkar, S., Hollis, R. J., Diekema, D. J. (2008). In Vitro Survey of Triazole Cross-Resistance among More than 700 Clinical Isolates of Aspergillus Species. J. Clin. Microbiol. 46: 2568-2572 [Abstract] [Full Text]  
  • Cowen, L. E., Steinbach, W. J. (2008). Stress, Drugs, and Evolution: the Role of Cellular Signaling in Fungal Drug Resistance. Eukaryot Cell 7: 747-764 [Full Text]  
  • Garcia-Effron, G., Dilger, A., Alcazar-Fuoli, L., Park, S., Mellado, E., Perlin, D. S. (2008). Rapid Detection of Triazole Antifungal Resistance in Aspergillus fumigatus. J. Clin. Microbiol. 46: 1200-1206 [Abstract] [Full Text]  
  • Mellado, E., Garcia-Effron, G., Alcazar-Fuoli, L., Melchers, W. J. G., Verweij, P. E., Cuenca-Estrella, M., Rodriguez-Tudela, J. L. (2007). A New Aspergillus fumigatus Resistance Mechanism Conferring In Vitro Cross-Resistance to Azole Antifungals Involves a Combination of cyp51A Alterations. Antimicrob. Agents Chemother. 51: 1897-1904 [Abstract] [Full Text]  
  • Patel, R., Mendrick, C., Knapp, C. C., Grist, R., McNicholas, P. M. (2007). Clinical Evaluation of the Sensititre YeastOne Plate for Testing Susceptibility of Filamentous Fungi to Posaconazole. J. Clin. Microbiol. 45: 2000-2001 [Abstract] [Full Text]  
  • Weiler, S., Bellmann, R., Kontoyiannis, D. P., Lewis, R. E., Krause, D. S., van Nieuwkoop, C., van Dissel, J. T., Ullmann, A. J., Chandrasekar, P., Patino, H., Cornely, O. A., Ullmann, A. J., Hardalo, C. (2007). Posaconazole Prophylaxis in Hematologic Cancer. NEJM 356: 2214-2218 [Full Text]  
  • Alcazar-Fuoli, L., Mellado, E., Garcia-Effron, G., Buitrago, M. J., Lopez, J. F., Grimalt, J. O., Cuenca-Estrella, J. M., Rodriguez-Tudela, J. L. (2006). Aspergillus fumigatus C-5 Sterol Desaturases Erg3A and Erg3B: Role in Sterol Biosynthesis and Antifungal Drug Susceptibility. Antimicrob. Agents Chemother. 50: 453-460 [Abstract] [Full Text]  
  • Garcia-Effron, G., Mellado, E., Gomez-Lopez, A., Alcazar-Fuoli, L., Cuenca-Estrella, M., Rodriguez-Tudela, J. L. (2005). Differences in Interactions between Azole Drugs Related to Modifications in the 14-{alpha} Sterol Demethylase Gene (cyp51A) of Aspergillus fumigatus. Antimicrob. Agents Chemother. 49: 2119-2121 [Abstract] [Full Text]  
  • Trama, J. P., Mordechai, E., Adelson, M. E. (2005). Detection of Aspergillus fumigatus and a Mutation That Confers Reduced Susceptibility to Itraconazole and Posaconazole by Real-Time PCR and Pyrosequencing. J. Clin. Microbiol. 43: 906-908 [Abstract] [Full Text]  
  • Balashov, S. V., Gardiner, R., Park, S., Perlin, D. S. (2005). Rapid, High-Throughput, Multiplex, Real-Time PCR for Identification of Mutations in the cyp51A Gene of Aspergillus fumigatus That Confer Resistance to Itraconazole. J. Clin. Microbiol. 43: 214-222 [Abstract] [Full Text]  
  • Chen, J., Li, H., Li, R., Bu, D., Wan, Z. (2005). Mutations in the cyp51A gene and susceptibility to itraconazole in Aspergillus fumigatus serially isolated from a patient with lung aspergilloma. J Antimicrob Chemother 55: 31-37 [Abstract] [Full Text]  
  • da Silva Ferreira, M. E., Capellaro, J. L., dos Reis Marques, E., Malavazi, I., Perlin, D., Park, S., Anderson, J. B., Colombo, A. L., Arthington-Skaggs, B. A., Goldman, M. H. S., Goldman, G. H. (2004). In Vitro Evolution of Itraconazole Resistance in Aspergillus fumigatus Involves Multiple Mechanisms of Resistance. Antimicrob. Agents Chemother. 48: 4405-4413 [Abstract] [Full Text]  
  • Mellado, E., Garcia-Effron, G., Alcazar-Fuoli, L., Cuenca-Estrella, M., Rodriguez-Tudela, J. L. (2004). Substitutions at Methionine 220 in the 14{alpha}-Sterol Demethylase (Cyp51A) of Aspergillus fumigatus Are Responsible for Resistance In Vitro to Azole Antifungal Drugs. Antimicrob. Agents Chemother. 48: 2747-2750 [Abstract] [Full Text]  
  • Xiao, L., Madison, V., Chau, A. S., Loebenberg, D., Palermo, R. E., McNicholas, P. M. (2004). Three-Dimensional Models of Wild-Type and Mutated Forms of Cytochrome P450 14{alpha}-Sterol Demethylases from Aspergillus fumigatus and Candida albicans Provide Insights into Posaconazole Binding. Antimicrob. Agents Chemother. 48: 568-574 [Abstract] [Full Text]  
  • Nascimento, A. M., Goldman, G. H., Park, S., Marras, S. A. E., Delmas, G., Oza, U., Lolans, K., Dudley, M. N., Mann, P. A., Perlin, D. S. (2003). Multiple Resistance Mechanisms among Aspergillus fumigatus Mutants with High-Level Resistance to Itraconazole. Antimicrob. Agents Chemother. 47: 1719-1726 [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»