Identification and characterization of a Cryptococcus neoformans ATP binding cassette (ABC) transporter-encoding gene, CnAFR1, involved in the resistance to fluconazole

Détails

ID Serval
serval:BIB_E34F9327D2A6
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Identification and characterization of a Cryptococcus neoformans ATP binding cassette (ABC) transporter-encoding gene, CnAFR1, involved in the resistance to fluconazole
Périodique
Molecular Microbiology
Auteur(s)
Posteraro  B., Sanguinetti  M., Sanglard  D., La Sorda  M., Boccia  S., Romano  L., Morace  G., Fadda  G.
ISSN
0950-382X (Print)
Statut éditorial
Publié
Date de publication
01/2003
Volume
47
Numéro
2
Pages
357-71
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jan
Résumé
Resistance to fluconazole is a possible event during prolonged suppressive drug therapy for cryptococ-cal meningitis, the most frequently encountered life-threatening manifestation of cryptococcosis. The knowledge of this resistance at the molecular level is important for management of cryptococcosis. In order to identify genes involved in azole resistance in Cryptococcus neoformans, a cDNA subtraction library technique was chosen as a strategy. First, a fluconazole-resistant mutant BPY22.17 was obtained from a susceptible clinical isolate BPY22 by in vitro exposure to the drug. Then, a subtractive hybridization procedure was used to compare gene expression between the obtained strains. We identified a cDNA overexpressed in the fluconazole-resistant strain BPY22.17 that was used as a probe to isolate the entire gene in a C. neoformans genomic library. Sequence analysis of this gene identified an ATP Binding Cassette (ABC) transporter-encoding gene called C. neoformans AntiFungal Resistance 1 (CnAFR1). Disruption of CnAFR1 gene in the resistant isolate (BPY22.17) resulted in an enhanced susceptibility of the knock-out mutant cnafr1 against fluconazole, whereas reintroduction of the gene in cnafr1 resulted in restoration of the resistance phenotype, thus confirming that CnAFR1 is involved in fluconazole resistance of C. neoformans. Our findings therefore reveal that an active drug efflux mechanism can be involved in the development of azole resistance in this important human pathogen.
Mots-clé
*ATP-Binding Cassette Transporters/chemistry/genetics/metabolism Amino Acid Sequence Antifungal Agents/*pharmacology Cloning, Molecular Cryptococcus neoformans/*drug effects/genetics/growth & development Drug Resistance, Fungal/*genetics Fluconazole/*pharmacology Fungal Proteins/chemistry/genetics/metabolism Gene Deletion Gene Library Humans Microbial Sensitivity Tests Molecular Sequence Data Nucleic Acid Hybridization/methods Sequence Alignment
Pubmed
Web of science
Open Access
Oui
Création de la notice
25/01/2008 14:40
Dernière modification de la notice
20/08/2019 16:07
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