title: Transcriptome analysis of Paracoccidioides brasiliensis cells undergoing mycelium-to-yeast transition.
abstract: Paracoccidioides brasiliensis is a thermodimorphic fungus associated with paracoccidioidomycosis (PCM), a systemic mycosis prevalent in South America. In humans, infection starts by inhalation of fungal propagules, which reach the pulmonary epithelium and transform into the yeast parasitic form. Thus, the mycelium-to-yeast transition is of particular interest because conversion to yeast is essential for infection. We have used a P. brasiliensis biochip carrying sequences of 4,692 genes from this fungus to monitor gene expression at several time points of the mycelium-to-yeast morphological shift (from 5 to 120 h). The results revealed a total of 2,583 genes that displayed statistically significant modulation in at least one experimental time point. Among the identified gene homologues, some encoded enzymes involved in amino acid catabolism, signal transduction, protein synthesis, cell wall metabolism, genome structure, oxidative stress response, growth control, and development. The expression pattern of 20 genes was independently verified by real-time reverse transcription-PCR, revealing a high degree of correlation between the data obtained with the two methodologies. One gene, encoding 4-hydroxyl-phenyl pyruvate dioxygenase (4-HPPD), was highly overexpressed during the mycelium-to-yeast differentiation, and the use of NTBC [2-(2-nitro-4-trifluoromethylbenzoyl)-cyclohexane-1,3-dione], a specific inhibitor of 4-HPPD activity, as well as that of NTBC derivatives, was able to inhibit growth and differentiation of the pathogenic yeast phase of the fungus in vitro. These data set the stage for further studies involving NTBC and its derivatives as new chemotherapeutic agents against PCM and confirm the potential of array-based approaches to identify new targets for the development of alternative treatments against pathogenic microorganisms.
authors.affiliation: Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Brazil.
authors.forename: Luiz R
authors.initials: LR
authors.lastname: Nunes
authors.forename: Regina
authors.initials: R
authors.lastname: Costa de Oliveira
authors.forename: Daniela Batista
authors.initials: DB
authors.lastname: Leite
authors.forename: Vivian Schmidt
authors.initials: VS
authors.lastname: da Silva
authors.forename: Everaldo
authors.initials: E
authors.lastname: dos Reis Marques
authors.forename: Márcia Eliana
authors.initials: ME
authors.lastname: da Silva Ferreira
authors.forename: Diógenes Custódio Duarte
authors.initials: DC
authors.lastname: Ribeiro
authors.forename: Luciano Angelo
authors.initials: LA
authors.lastname: de Souza Bernardes
authors.forename: Maria Helena S
authors.initials: MH
authors.lastname: Goldman
authors.forename: Rosana
authors.initials: R
authors.lastname: Puccia
authors.forename: Luiz R
authors.initials: LR
authors.lastname: Travassos
authors.forename: Wagner L
authors.initials: WL
authors.lastname: Batista
authors.forename: Marina Pasetto
authors.initials: MP
authors.lastname: Nóbrega
authors.forename: Francisco G
authors.initials: FG
authors.lastname: Nobrega
authors.forename: Ding-Yah
authors.initials: DY
authors.lastname: Yang
authors.forename: Carlos A
authors.initials: CA
authors.lastname: de Bragança Pereira
authors.forename: Gustavo H
authors.initials: GH
authors.lastname: Goldman
mesh_terms: D015111:4-Hydroxyphenylpyruvate Dioxygenase / Q000037:antagonists & inhibitors; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003470:Culture Media; D003512:Cyclohexanones / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D006801:Humans; D046228:Microarray Analysis; D015394:Molecular Structure; D025282:Mycelium / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D009579:Nitrobenzoates / Q000494:pharmacology; D010228:Paracoccidioides / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D010229:Paracoccidioidomycosis / Q000209:etiology; D013696:Temperature; D014158:Transcription, Genetic; D015003:Yeasts / Q000166:cytology* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
chemical_list: D003470:Culture Media; D003512:Cyclohexanones; D004791:Enzyme Inhibitors; D009579:Nitrobenzoates; D015111:4-Hydroxyphenylpyruvate Dioxygenase; C077073:nitisinone