title: Identification of novel genes conferring altered azole susceptibility in  Aspergillus fumigatus.
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.
authors.affiliation: Manchester Academic Health Science Centre, NIHR Translational Research Facility in Respiratory Medicine, University Hospital of South Manchester NHS Foundation Trust, The University of Manchester, Manchester, UK. paul.bowyer@manchester.ac.uk
authors.forename: Paul
authors.initials: P
authors.lastname: Bowyer
authors.forename: Juan
authors.initials: J
authors.lastname: Mosquera
authors.forename: Michael
authors.initials: M
authors.lastname: Anderson
authors.forename: Mike
authors.initials: M
authors.lastname: Birch
authors.forename: Michael
authors.initials: M
authors.lastname: Bromley
authors.forename: David W
authors.initials: DW
authors.lastname: Denning
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D001232:Aspergillus fumigatus / Q000187:drug effects* / Q000235:genetics*; D015139:Blotting, Southern; D002874:Chromosome Mapping; D025141:Drug Resistance, Fungal; D005800:Genes, Fungal; D017964:Itraconazole / Q000494:pharmacology*; D016254:Mutagenesis, Insertional; D010641:Phenotype; D010957:Plasmids
chemical_list: D000935:Antifungal Agents; D017964:Itraconazole