Identification of Novel Genes Conferring Altered Azole Susceptibility in Aspergillus Fumigatus

Overview

Journal FEMS Microbiol Lett

Publisher Oxford University Press

Specialty Microbiology

Date 2012 Apr 19

PMID 22509997

Citations 19

Authors

Paul Bowyer

Juan Mosquera

Michael Anderson

Mike Birch

Michael Bromley

David W Denning

Affiliations

Soon will be listed here.

Abstract

Azoles are currently the mainstay of antifungal treatment both in agricultural and in clinical settings. Although the target site of azole action is well studied, the basis of azole resistance and the ultimate mode of action of the drug in fungi are poorly understood. To gain a deeper insight into these aspects of azole action, restriction-mediated plasmid integration (REMI) was used to create azole sensitive and resistant strains of the clinically important fungus Aspergillus fumigatus. Four azole sensitive insertions and four azole-resistant insertions were characterized. Three phenotypes could be re-created in wild-type AF210 by reintegration of rescued plasmid and a further four could be confirmed by complementation of the mutant phenotype with a copy of the wild-type gene predicted to be disrupted by the original insertional event. Six insertions were in genes not previously associated with azole sensitivity or resistance. Two insertions occur in transporter genes that may affect drug efflux, whereas others may affect transcriptional regulation of sterol biosynthesis genes and NADH metabolism in the mitochondrion. Two insertions are in genes of unknown function.

Citing Articles

A highly multiplexed assay to monitor pathogenicity, fungicide resistance and gene flow in the fungal wheat pathogen Zymoseptoria tritici.

Bellah H, Gazeau G, Gelisse S, Amezrou R, Marcel T, Croll D PLoS One. 2023; 18(2):e0281181.

PMID: 36745583 PMC: 9901794. DOI: 10.1371/journal.pone.0281181.

Genomic Data Mining Reveals Abundant Uncharacterized Transporters in and .

Cai H, Zhang H, Guo D, Wang Y, Gu J J Fungi (Basel). 2022; 8(10).

PMID: 36294626 PMC: 9604845. DOI: 10.3390/jof8101064.

The Anti-Fungal Activity of Nitropropenyl Benzodioxole (NPBD), a Redox-Thiol Oxidant and Tyrosine Phosphatase Inhibitor.

Nicoletti G, White K Antibiotics (Basel). 2022; 11(9).

PMID: 36139967 PMC: 9495065. DOI: 10.3390/antibiotics11091188.

Genome-Wide Association Analysis for Triazole Resistance in .

Fan Y, Wang Y, Korfanty G, Archer M, Xu J Pathogens. 2021; 10(6).

PMID: 34199862 PMC: 8227032. DOI: 10.3390/pathogens10060701.

Identification of Novel Mutations Contributing to Azole Tolerance of Aspergillus fumigatus through Exposure to Tebuconazole.

Toyotome T, Onishi K, Sato M, Kusuya Y, Hagiwara D, Watanabe A Antimicrob Agents Chemother. 2021; 65(9):e0265720.

PMID: 34125587 PMC: 8370235. DOI: 10.1128/AAC.02657-20.

References

Mellado E, Garcia-Effron G, Alcazar-Fuoli L, Cuenca-Estrella M, Rodriguez-Tudela J . Substitutions at methionine 220 in the 14alpha-sterol demethylase (Cyp51A) of Aspergillus fumigatus are responsible for resistance in vitro to azole antifungal drugs. Antimicrob Agents Chemother. 2004; 48(7):2747-50. PMC: 434164. DOI: 10.1128/AAC.48.7.2747-2750.2004. View

Howard S, Cerar D, Anderson M, Albarrag A, Fisher M, Pasqualotto A . Frequency and evolution of Azole resistance in Aspergillus fumigatus associated with treatment failure. Emerg Infect Dis. 2009; 15(7):1068-76. PMC: 2744247. DOI: 10.3201/eid1507.090043. View

Latge J, Calderone R . Host-microbe interactions: fungi invasive human fungal opportunistic infections. Curr Opin Microbiol. 2002; 5(4):355-8. DOI: 10.1016/s1369-5274(02)00343-0. View

Sanglard D, Ischer F, Monod M, Bille J . Cloning of Candida albicans genes conferring resistance to azole antifungal agents: characterization of CDR2, a new multidrug ABC transporter gene. Microbiology (Reading). 1997; 143 ( Pt 2):405-416. DOI: 10.1099/00221287-143-2-405. View

Mellado E, Garcia-Effron G, Buitrago M, Alcazar-Fuoli L, Cuenca-Estrella M, Rodriguez-Tudela J . Targeted gene disruption of the 14-alpha sterol demethylase (cyp51A) in Aspergillus fumigatus and its role in azole drug susceptibility. Antimicrob Agents Chemother. 2005; 49(6):2536-8. PMC: 1140498. DOI: 10.1128/AAC.49.6.2536-2538.2005. View

Chandrasekar P, Cutright J, Manavathu E . Aspergillus: rising frequency of clinical isolation and continued susceptibility to antifungal agents, 1994-1999. Diagn Microbiol Infect Dis. 2002; 41(4):211-4. DOI: 10.1016/s0732-8893(01)00295-4. View

Lu S, Lyngholm L, Yang G, Bronson C, Yoder O, Turgeon B . Tagged mutations at the Tox1 locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration. Proc Natl Acad Sci U S A. 1994; 91(26):12649-53. PMC: 45496. DOI: 10.1073/pnas.91.26.12649. View

Bolker M, Bohnert H, Braun K, Gorl J, Kahmann R . Tagging pathogenicity genes in Ustilago maydis by restriction enzyme-mediated integration (REMI). Mol Gen Genet. 1995; 248(5):547-52. DOI: 10.1007/BF02423450. View

Yoshida Y, Aoyama Y . Yeast cytochrome P-450 catalyzing lanosterol 14 alpha-demethylation. I. Purification and spectral properties. J Biol Chem. 1984; 259(3):1655-60. View

10.

Nierman W, Pain A, Anderson M, Wortman J, Kim H, Arroyo J . Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature. 2005; 438(7071):1151-6. DOI: 10.1038/nature04332. View

11.

Nascimento A, Goldman G, Park S, Marras S, Delmas G, Oza U . Multiple resistance mechanisms among Aspergillus fumigatus mutants with high-level resistance to itraconazole. Antimicrob Agents Chemother. 2003; 47(5):1719-26. PMC: 153329. DOI: 10.1128/AAC.47.5.1719-1726.2003. View

12.

Marques I, Duarte M, Assuncao J, Ushakova A, Videira A . Composition of complex I from Neurospora crassa and disruption of two "accessory" subunits. Biochim Biophys Acta. 2005; 1707(2-3):211-20. DOI: 10.1016/j.bbabio.2004.12.003. View

13.

Marichal P, Koymans L, Willemsens S, Bellens D, Verhasselt P, Luyten W . Contribution of mutations in the cytochrome P450 14alpha-demethylase (Erg11p, Cyp51p) to azole resistance in Candida albicans. Microbiology (Reading). 1999; 145 ( Pt 10):2701-2713. DOI: 10.1099/00221287-145-10-2701. View

14.

Denning D, Venkateswarlu K, Oakley K, Anderson M, Manning N, Stevens D . Itraconazole resistance in Aspergillus fumigatus. Antimicrob Agents Chemother. 1997; 41(6):1364-8. PMC: 163916. DOI: 10.1128/AAC.41.6.1364. View

15.

Arunmozhi Balajee S, Weaver M, Imhof A, Gribskov J, Marr K . Aspergillus fumigatus variant with decreased susceptibility to multiple antifungals. Antimicrob Agents Chemother. 2004; 48(4):1197-203. PMC: 375298. DOI: 10.1128/AAC.48.4.1197-1203.2004. View

16.

Chen J, Li H, Li R, Bu D, Wan Z . Mutations in the cyp51A gene and susceptibility to itraconazole in Aspergillus fumigatus serially isolated from a patient with lung aspergilloma. J Antimicrob Chemother. 2004; 55(1):31-7. DOI: 10.1093/jac/dkh507. View

17.

Schaller M, Schafer W, Korting H, Hube B . Differential expression of secreted aspartyl proteinases in a model of human oral candidosis and in patient samples from the oral cavity. Mol Microbiol. 1998; 29(2):605-15. DOI: 10.1046/j.1365-2958.1998.00957.x. View

18.

Verweij P, Mensink M, Rijs A, Donnelly J, Meis J, Denning D . In-vitro activities of amphotericin B, itraconazole and voriconazole against 150 clinical and environmental Aspergillus fumigatus isolates. J Antimicrob Chemother. 1998; 42(3):389-92. DOI: 10.1093/jac/42.3.389. View

19.

Vogeser M, Wanders A, Haas A, Ruckdeschel G . A four-year review of fatal Aspergillosis. Eur J Clin Microbiol Infect Dis. 1999; 18(1):42-5. DOI: 10.1007/s100960050224. View

20.

Wright G, Parent T, Honek J . Non-sterol structural probes of the lanosterol 14 alpha-demethylase from Saccharomyces cerevisiae. Biochim Biophys Acta. 1990; 1040(1):95-101. DOI: 10.1016/0167-4838(90)90151-5. View