title: The Aspergillus fumigatus siderophore biosynthetic gene sidA, encoding L-ornithine N5-oxygenase, is required for virulence.
abstract: Aspergillus fumigatus is the leading cause of invasive mold infection and is a serious problem in immunocompromised populations worldwide. We have previously shown that survival of A. fumigatus in serum may be related to secretion of siderophores. In this study, we identified and characterized the sidA gene of A. fumigatus, which encodes l-ornithine N(5)-oxygenase, the first committed step in hydroxamate siderophore biosynthesis. A. fumigatus sidA codes for a protein of 501 amino acids with significant homology to other fungal l-ornithine N(5)-oxygenases. A stable DeltasidA strain was created by deletion of A. fumigatus sidA. This strain was unable to synthesize the siderophores N',N",N'''-triacetylfusarinine C (TAF) and ferricrocin. Growth of the DeltasidA strain was the same as that of the wild type in rich media; however, the DeltasidA strain was unable to grow in low-iron defined media or media containing 10% human serum unless supplemented with TAF or ferricrocin. No significant differences in ferric reduction activities were observed between the parental strain and the DeltasidA strain, indicating that blocking siderophore secretion did not result in upregulation of this pathway. Unlike the parental strain, the DeltasidA strain was unable to remove iron from human transferrin. A rescued strain (DeltasidA + sidA) was constructed; it produced siderophores and had the same growth as the wild type on iron-limited media. Unlike the wild-type and rescued strains, the DeltasidA strain was avirulent in a mouse model of invasive aspergillosis, indicating that sidA is necessary for A. fumigatus virulence.
authors.affiliation: Department of Biological Sciences, Simon Fraser University, 8888 University Drive, Burnaby, British Columbia V5A 1S6, Canada. mmoore@sfu.ca.
authors.forename: Anna H T
authors.initials: AH
authors.lastname: Hissen
authors.forename: Adrian N C
authors.initials: AN
authors.lastname: Wan
authors.forename: Mark L
authors.initials: ML
authors.lastname: Warwas
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Pinto
authors.forename: Margo M
authors.initials: MM
authors.lastname: Moore
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001228:Aspergillosis / Q000201:enzymology / Q000382:microbiology* / Q000473:pathology; D001232:Aspergillus fumigatus / Q000235:genetics* / Q000472:pathogenicity*; D004195:Disease Models, Animal; D005291:Ferrichrome / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000378:metabolism; D008168:Lung / Q000382:microbiology / Q000473:pathology; D008172:Lung Diseases, Fungal / Q000201:enzymology / Q000382:microbiology* / Q000473:pathology; D051379:Mice; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D016415:Sequence Alignment; D017262:Siderophores / Q000096:biosynthesis*; D014774:Virulence
chemical_list: D006877:Hydroxamic Acids; D017262:Siderophores; C051730:fusarinine; D005291:Ferrichrome; C021678:ferricrocin; D006899:Mixed Function Oxygenases; C059419:ornithine N5-oxygenase