Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole

Patrícia Fernanda Zambuzzi-Carvalho, Amanda Gregorim Fernandes, Marize Campos Valadares, Patrícia De Mello Tavares, Joshua D. Nosanchuk, Célia Maria De Almeida Soares, Maristela Pereira

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)477-492
Number of pages16
JournalMedical Mycology
Volume53
Issue number5
DOIs
StatePublished - 2015

Fingerprint

animal pathogenic fungi
Paracoccidioides
Sulfamethoxazole
sulfamethoxazole
Fungi
Paracoccidioidomycosis
Mycoses
sulfonamides
Sulfonamides
drugs
Aminobenzoates
Dihydropteroate Synthase
aminobenzoic acids
Pharmaceutical Preparations
Tetrazolium Salts
Genes
4-Aminobenzoic Acid
genes
Latin America
mycoses

Keywords

  • Antifungal
  • Paracoccidioides lutzii
  • Representational difference analysis
  • Sulfamethoxazole
  • Transcriptome

ASJC Scopus subject areas

  • veterinary(all)
  • Infectious Diseases

Cite this

Zambuzzi-Carvalho, P. F., Fernandes, A. G., Valadares, M. C., De Mello Tavares, P., Nosanchuk, J. D., De Almeida Soares, C. M., & Pereira, M. (2015). Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. Medical Mycology, 53(5), 477-492. https://doi.org/10.1093/mmy/myv011

Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. / Zambuzzi-Carvalho, Patrícia Fernanda; Fernandes, Amanda Gregorim; Valadares, Marize Campos; De Mello Tavares, Patrícia; Nosanchuk, Joshua D.; De Almeida Soares, Célia Maria; Pereira, Maristela.

In: Medical Mycology, Vol. 53, No. 5, 2015, p. 477-492.

Research output: Contribution to journalArticle

Zambuzzi-Carvalho, PF, Fernandes, AG, Valadares, MC, De Mello Tavares, P, Nosanchuk, JD, De Almeida Soares, CM & Pereira, M 2015, 'Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole', Medical Mycology, vol. 53, no. 5, pp. 477-492. https://doi.org/10.1093/mmy/myv011
Zambuzzi-Carvalho PF, Fernandes AG, Valadares MC, De Mello Tavares P, Nosanchuk JD, De Almeida Soares CM et al. Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. Medical Mycology. 2015;53(5):477-492. https://doi.org/10.1093/mmy/myv011
Zambuzzi-Carvalho, Patrícia Fernanda ; Fernandes, Amanda Gregorim ; Valadares, Marize Campos ; De Mello Tavares, Patrícia ; Nosanchuk, Joshua D. ; De Almeida Soares, Célia Maria ; Pereira, Maristela. / Transcriptional profile of the human pathogenic fungus Paracoccidioides lutzii in response to sulfamethoxazole. In: Medical Mycology. 2015 ; Vol. 53, No. 5. pp. 477-492.
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