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Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole.
Med Mycol. 2015 Jun; 53(5):477-92.MM

Abstract

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.

Authors+Show Affiliations

Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, GO, Brazil.Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, GO, Brazil.Laboratório de Farmacologia e Toxicologia Celular, Faculdade de Farmácia, Universidade Federal de Goiás, Goiânia, GO, Brazil.Departments of Medicine and Microbiology and Immunology, Albert Einstein College of Medicine, New York, NY, USA.Departments of Medicine and Microbiology and Immunology, Albert Einstein College of Medicine, New York, NY, USA.Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, GO, Brazil.Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, GO, Brazil maristelaufg@gmail.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25850856

Citation

Zambuzzi-Carvalho, Patrícia Fernanda, et al. "Transcriptional Profile of the Human Pathogenic Fungus Paracoccidioides Lutzii in Response to Sulfamethoxazole." Medical Mycology, vol. 53, no. 5, 2015, pp. 477-92.
Zambuzzi-Carvalho PF, Fernandes AG, Valadares MC, et al. Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. Med Mycol. 2015;53(5):477-92.
Zambuzzi-Carvalho, P. F., Fernandes, A. G., Valadares, M. C., Tavares, P. d. e. . M., Nosanchuk, J. D., Soares, C. M., & Pereira, M. (2015). Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. Medical Mycology, 53(5), 477-92. https://doi.org/10.1093/mmy/myv011
Zambuzzi-Carvalho PF, et al. Transcriptional Profile of the Human Pathogenic Fungus Paracoccidioides Lutzii in Response to Sulfamethoxazole. Med Mycol. 2015;53(5):477-92. PubMed PMID: 25850856.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. AU - Zambuzzi-Carvalho,Patrícia Fernanda, AU - Fernandes,Amanda Gregorim, AU - Valadares,Marize Campos, AU - Tavares,Patrícia de Mello, AU - Nosanchuk,Joshua D, AU - Soares,Célia Maria de Almeida, AU - Pereira,Maristela, Y1 - 2015/04/07/ PY - 2015/01/27/accepted PY - 2014/10/15/received PY - 2015/4/9/entrez PY - 2015/4/9/pubmed PY - 2016/3/15/medline KW - Paracoccidioides lutzii KW - antifungal KW - representational difference analysis KW - sulfamethoxazole KW - transcriptome SP - 477 EP - 92 JF - Medical mycology JO - Med. Mycol. VL - 53 IS - 5 N2 - 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. SN - 1460-2709 UR - https://www.unboundmedicine.com/medline/citation/25850856/Transcriptional_profile_of_the_human_pathogenic_fungus_Paracoccidioides_lutzii_in_response_to_sulfamethoxazole_ L2 - https://academic.oup.com/mmy/article-lookup/doi/10.1093/mmy/myv011 DB - PRIME DP - Unbound Medicine ER -