Open Access Research article

Transcriptional profile of Paracoccidioides induced by oenothein B, a potential antifungal agent from the Brazilian Cerrado plant Eugenia uniflora

Patrícia Fernanda Zambuzzi-Carvalho1, Patrícia Kott Tomazett1, Suzana Costa Santos2, Pedro Henrique Ferri2, Clayton Luiz Borges1, Wellington Santos Martins3, Célia Maria de Almeida Soares1 and Maristela Pereira1*

Author Affiliations

1 Departamento de Bioquímica e Biologia Molecular, Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICBII, Campus II, Universidade Federal de Goiás, C.P. 131, 74001-970 Goiânia, GO, Brazil

2 Laboratório de Bioatividade Molecular, Instituto de Química, Universidade Federal de Goiás, Goiânia, GO, Brazil

3 Instituto de Informática, Universidade Federal de Goiás, Goiânia, Goiás, Brazil

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BMC Microbiology 2013, 13:227  doi:10.1186/1471-2180-13-227

Published: 12 October 2013

Abstract

Background

The compound oenothein B (OenB), which is isolated from the leaves of Eugenia uniflora, a Brazilian Cerrado plant, interferes with Paracoccidioides yeast cell morphology and inhibits 1,3-β-D-glucan synthase (PbFKS1) transcript accumulation, which is involved in cell wall synthesis. In this work we examined the gene expression changes in Paracoccidioides yeast cells following OenB treatment in order to investigate the adaptive cellular responses to drug stress.

Results

We constructed differential gene expression libraries using Representational Difference Analysis (RDA) of Paracoccidioides yeast cells treated with OenB for 90 and 180 min. Treatment for 90 min resulted in the identification of 463 up-regulated expressed sequences tags (ESTs) and 104 down-regulated ESTs. For the 180 min treatment 301 up-regulated ESTs and 143 down-regulated were identified. Genes involved in the cell wall biosynthesis, such as GLN1, KRE6 and FKS1, were found to be regulated by OenB. Infection experiments in macrophages corroborated the in vitro results. Fluorescence microscopy showed increased levels of chitin in cells treated with OenB. The carbohydrate polymer content of the cell wall of the fungus was also evaluated, and the results corroborated with the transcriptional data. Several other genes, such as those involved in a variety of important cellular processes (i.e., membrane maintenance, stress and virulence) were found to be up-regulated in response to OenB treatment.

Conclusions

The exposure of Paracoccidioides to OenB resulted in a complex altered gene expression profile. Some of the changes may represent specific adaptive responses to this compound in this important pathogenic fungus.

Keywords:
Paracoccidioides; Antifungal; Oenothein B; Transcriptome; Cell wall