title: Identification and characterization of an SKN7 homologue in Cryptococcus neoformans.
abstract: Cryptococcus neoformans is an encapsulated fungal pathogen that primarily infects the central nervous system of immunocompromised individuals, causing life-threatening meningoencephalitis. The capacity of C. neoformans to subvert host defenses and disseminate by intracellular parasitism of alveolar macrophages in the immune-compromised host has led to studies to evaluate genes associated with C. neoformans resistance to oxidative stress. In the present study, we identify and characterize a C. neoformans homologue to SKN7, a transcription factor in Saccharomyces cerevisiae that regulates the oxidative stress response, cell cycle, and cell wall biosynthesis. To examine the contribution of SKN7 in the pathogenesis of fungal infections, we created skn7 mutants via targeted disruption. The skn7 mutants were observed to be more susceptible to reactive oxygen species in vitro and were significantly less virulent than the wild-type strain and a reconstituted strain as measured by cumulative survival in the mouse inhalational model. The Skn7 protein was observed to be important for expression of thioredoxin reductase in response to oxidative challenge. Interestingly, skn7 mutants were also observed to flocculate following in vitro culture, a novel phenotype not observed in skn7 mutants derived from other fungi. These findings demonstrate that SKN7 contributes to the virulence composite but is not required for pathogenicity in C. neoformans. In addition, flocculation of C. neoformans skn7 mutants suggests a potentially unique function of SKN7 not previously observed in other cryptococcal strains or skn7 mutants.
authors.affiliation: Department of Medicine/Division of Infectious Diseases, Duke University Medical Center, Duke South, Stead Bldg., Box 3353, Durham, NC 27710, USA. floyd.wormley@duke.edu
authors.forename: F L
authors.initials: FL
authors.lastname: Wormley
authors.forename: G
authors.initials: G
authors.lastname: Heinrich
authors.forename: J L
authors.initials: JL
authors.lastname: Miller
authors.forename: J R
authors.initials: JR
authors.lastname: Perfect
authors.forename: G M
authors.initials: GM
authors.lastname: Cox
mesh_terms: D000818:Animals; D003453:Cryptococcosis / Q000235:genetics / Q000378:metabolism; D003455:Cryptococcus neoformans / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D005427:Flocculation; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D008171:Lung Diseases / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D030781:Organisms, Genetically Modified; D018384:Oxidative Stress / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014774:Virulence; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
chemical_list: D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C503125:SKN7 protein, Cryptococcus neoformans; C083962:SKN7 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D020122:tert-Butylhydroperoxide