Transcriptome analysis of Paracoccidioides brasiliensis cells undergoing mycelium-to-yeast transition.
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.
Nunes
Luiz R
LR
Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Brazil.
Costa de Oliveira
Regina
R
Leite
Daniela Batista
DB
da Silva
Vivian Schmidt
VS
dos Reis Marques
Everaldo
E
da Silva Ferreira
Márcia Eliana
ME
Ribeiro
Diógenes Custódio Duarte
DC
de Souza Bernardes
Luciano Angelo
LA
Goldman
Maria Helena S
MH
Puccia
Rosana
R
Travassos
Luiz R
LR
Batista
Wagner L
WL
Nóbrega
Marina Pasetto
MP
Nobrega
Francisco G
FG
Yang
Ding-Yah
DY
de Bragança Pereira
Carlos A
CA
Goldman
Gustavo H
GH
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
D003470:Culture Media; D003512:Cyclohexanones; D004791:Enzyme Inhibitors; D009579:Nitrobenzoates; D015111:4-Hydroxyphenylpyruvate Dioxygenase; C077073:nitisinone