Thiol peroxidase is critical for virulence and resistance to nitric oxide and peroxide in the fungal pathogen, Cryptococcus neoformans.
Cryptococcus neoformans is a fungal pathogen most commonly causing meningitis in immunocompromised patients. Current therapies are inadequate, and novel antifungal targets are needed. We have identified by proteomics two thiol peroxidases that are differentially expressed at 37 degrees C, the temperature of the mammalian host. Consistent with their antioxidant role, we show that the genes encoding these thiol-specific antioxidants, TSA1 and TSA3, are transcriptionally induced when C. neoformans is exposed to hydrogen peroxide. Genome sequence analysis of C. neoformans revealed a third thiol peroxidase, TSA4. We constructed single, double and triple mutants of the thiol peroxidase genes through homologous recombination and analysed their function by comparing the growth of these mutants with that of the wild-type strain. The tsa1 Delta mutant shows sensitivity to hydrogen peroxide and t-butylhydroperoxide, as well as significant growth retardation at 25 degrees C and 38.5 degrees C. The tsa1 Delta mutant is also sensitive to NO, demonstrating a link between oxidative and nitrosative stress pathways. In two mouse models of cryptococcosis, the tsa1 Delta mutant is significantly less virulent.
Missall
Tricia Ann
TA
Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, 1402 S Grand Boulevard, Saint Louis, MO 63104, USA.
Pusateri
Mary Ellen
ME
Lodge
Jennifer K
JK
D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D003455:Cryptococcus neoformans / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D051379:Mice; D008808:Mice, Inbred CBA; D009569:Nitric Oxide / Q000494:pharmacology*; D016877:Oxidants / Q000378:metabolism; D010544:Peroxidases / Q000235:genetics / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013696:Temperature; D014774:Virulence; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
D000975:Antioxidants; D005656:Fungal Proteins; D016877:Oxidants; D013438:Sulfhydryl Compounds; D009569:Nitric Oxide; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide; D010544:Peroxidases