title: Transcriptional profile of Paracoccidioides induced by oenothein B, a potential antifungal agent from the Brazilian Cerrado plant Eugenia uniflora.
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.
authors.affiliation: 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. maristelaufg@gmail.com.
authors.forename: Patrícia Fernanda
authors.initials: PF
authors.lastname: Zambuzzi-Carvalho
authors.forename: Patrícia Kott
authors.initials: PK
authors.lastname: Tomazett
authors.forename: Suzana Costa
authors.initials: SC
authors.lastname: Santos
authors.forename: Pedro Henrique
authors.initials: PH
authors.lastname: Ferri
authors.forename: Clayton Luiz
authors.initials: CL
authors.lastname: Borges
authors.forename: Wellington Santos
authors.initials: WS
authors.lastname: Martins
authors.forename: Célia Maria
authors.initials: CM
authors.lastname: de Almeida Soares
authors.forename: Maristela
authors.initials: M
authors.lastname: Pereira
mesh_terms: D000935:Antifungal Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001938:Brazil; D020869:Gene Expression Profiling; D047348:Hydrolyzable Tannins / Q000302:isolation & purification / Q000494:pharmacology*; D010228:Paracoccidioides / Q000187:drug effects* / Q000235:genetics; D027842:Syzygium / Q000737:chemistry
chemical_list: D000935:Antifungal Agents; D047348:Hydrolyzable Tannins; C080077:oenothein B