Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole
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Data
2015
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Resumo
Paracoccidioidomycosis (PCM) is the most prevalent mycosis in Latin America and is
caused by a group of fungi within the Paracoccidioides genus. The disease may present
clinical and pathological manifestations ranging from asymptomatic pneumonia pulmonary
lesions, to disseminated forms involving multiple organs. Sulfonamides were
the first drugs used to treat PCM and are still used against this fungal infection. Sulfa
drugs are competitive antagonists of ρ-aminobenzoic acid (PABA), a reaction catalyzed
by dihydropteroate synthase (DHPS). However, the molecular effects of sulfonamides
against the Paracoccidioides genus are unknown. The aim of this work was to investigate
the global mechanism of action of sulfamethoxazole on Paracoccidioides lutzii. Yeast
cells were grown on minimum medium in the presence or absence of sulfamethoxazole
to construct EST libraries. The representational difference analysis (RDA) technique
was used to identify up- and down-regulated P. lutzii genes after treatment with sulfamethoxazole.
Approximately six transcripts related to mitochondrial function were differentially
expressed. To confirm the RDA and bioinformatics results, several relevant
genes were studied with quantitative real-time polymerase chain reaction (qRT-PCR) to
evaluate their levels of expression. To confirm the impact of sulfamethoxazole on mitochondria,
we measured the reduction of tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide (MTT) by P. lutzii with or without exposure to the drug.
MTT assays reveal that sulfamethoxazole produces a marked dose-dependent adverse effect on P. lutzii. The transcriptional activity of selected genes in infected macrophages
corroborated our in vitro results. The results indicated that sulfamethoxazole acts in
P. lutzii as a competitor for amino acid, nucleic acids and folate cofactor biosynthesis,
disrupting mitochondrial functions.
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Palavras-chave
Paracoccidioides lutzii, Antifungal, Sulfamethoxazole, Transcriptome, Representational difference analysis
Citação
ZAMBUZZI-CARVALHO, Patrícia Fernanda et al. Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. Medical Mycology, Oxford, v. 53, n. 5, p. 477-492, 2015.