title: Identification and Analysis of the Role of Superoxide Dismutases Isoforms in the Pathogenesis of Paracoccidioides spp.
abstract: The ability of Paracoccidioides to defend itself against reactive oxygen species (ROS) produced by host effector cells is a prerequisite to survive. To counteract these radicals, Paracoccidioides expresses, among different antioxidant enzymes, superoxide dismutases (SODs). In this study, we identified six SODs isoforms encoded by the Paracoccidioides genome. We determined gene expression levels of representative isolates of the phylogenetic lineages of Paracoccidioides spp. (S1, PS2, PS3 and Pb01-like) using quantitative RT-PCR. Assays were carried out to analyze SOD gene expression of yeast cells, mycelia cells, the mycelia-to-yeast transition and the yeast-to-mycelia germination, as well as under treatment with oxidative agents and during interaction with phagocytic cells. We observed an increased expression of PbSOD1 and PbSOD3 during the transition process, exposure to oxidative agents and interaction with phagocytic cells, suggesting that these proteins could assist in combating the superoxide radicals generated during the host-pathogen interaction. Using PbSOD1 and PbSOD3 knockdown strains we showed these genes are involved in the response of the fungus against host effector cells, particularly the oxidative stress response, and in a mouse model of infection. Protein sequence analysis together with functional analysis of knockdown strains seem to suggest that PbSOD3 expression is linked with a pronounced extracellular activity while PbSOD1 seems more related to intracellular requirements of the fungus. Altogether, our data suggests that P. brasiliensis actively responds to the radicals generated endogenously during metabolism and counteracts the oxidative burst of immune cells by inducing the expression of SOD isoforms.
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.|Institute of Biology, Universidad de Antioquia, Medellín, Colombia.
authors.forename: Diana
authors.initials: D
authors.lastname: Tamayo
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.|Institute of Biology, Universidad de Antioquia, Medellín, Colombia.
authors.forename: José F
authors.initials: JF
authors.lastname: Muñoz
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.|Institute of Biology, Universidad de Antioquia, Medellín, Colombia.
authors.forename: Ángela
authors.initials: Á
authors.lastname: Lopez
authors.affiliation: Institute of Biomedical Science; Department of Microbiology; Universidade do São Paulo; São Paulo, Brazil.
authors.forename: Martha
authors.initials: M
authors.lastname: Urán
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.
authors.forename: Juan
authors.initials: J
authors.lastname: Herrera
authors.affiliation: Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICBII, Goiânia, Brazil.
authors.forename: Clayton L
authors.initials: CL
authors.lastname: Borges
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.
authors.forename: Ángela
authors.initials: Á
authors.lastname: Restrepo
authors.affiliation: Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICBII, Goiânia, Brazil.
authors.forename: Celia M
authors.initials: CM
authors.lastname: Soares
authors.affiliation: Institute of Biomedical Science; Department of Microbiology; Universidade do São Paulo; São Paulo, Brazil.
authors.forename: Carlos P
authors.initials: CP
authors.lastname: Taborda
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.|Department of Biological Sciences, School of Sciences, Universidad Eafit, Medellín, Colombia.
authors.forename: Agostinho J
authors.initials: AJ
authors.lastname: Almeida
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.|School of Medicine, Universidad de Antioquia, Medellín, Colombia.
authors.forename: Juan G
authors.initials: JG
authors.lastname: McEwen
authors.affiliation: Cellular and Molecular Biology Unit, Corporación para Investigaciones Biológicas, Medellín, Colombia.|MICROBA research group, School of Microbiology, Universidad de Antioquia, Medellín, Colombia.
authors.forename: Orville
authors.initials: O
authors.lastname: Hernández
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D008297:Male; D008807:Mice, Inbred BALB C; D010228:Paracoccidioides / Q000201:enzymology* / Q000472:pathogenicity*; D010229:Paracoccidioidomycosis / Q000473:pathology*; D020033:Protein Isoforms / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000378:metabolism*
chemical_list: D020033:Protein Isoforms; D013482:Superoxide Dismutase