Identification of genes preferentially expressed in the pathogenic yeast phase of Paracoccidioides brasiliensis, using suppression subtraction hybridization and differential macroarray analysis

Identification of genes preferentially expressed in the pathogenic yeast phase of Paracoccidioides brasiliensis, using suppression subtraction hybridization and differential macroarray analysis

Autor Marques, E. R. Google Scholar
Ferreira, MES Google Scholar
Drummond, R. D. Google Scholar
Felix, J. M. Google Scholar
Menossi, M. Google Scholar
Savoldi, M. Google Scholar
Travassos, L. R. Google Scholar
Puccia, R. Google Scholar
Batista, W. L. Google Scholar
Carvalho, K. C. Google Scholar
Goldman, MHS Google Scholar
Goldman, G. H. Google Scholar
Instituição Universidade de São Paulo (USP)
Universidade Estadual de Campinas (UNICAMP)
Universidade Federal de São Paulo (UNIFESP)
Resumo Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America. Pathogenicity appears to be intimately related to the dimorphic transition from the hyphal to the yeast form, which is induced by a shift from environmental temperature to the temperature of the mammalian host. Little information is available on the P. brasiliensis genes that are necessary during the pathogenic phase. We have therefore undertaken Suppression Subtraction Hybridization (SSH) and macroarray analyses with the aim of identifying genes that are preferentially expressed in the yeast phase. Genes identified by both procedures as being more highly expressed in the yeast phase are involved in basic metabolism, signal transduction, growth and morphogenesis, and sulfur metabolism. in order to test whether the observed changes in gene expression reflect the differences between the growth conditions used to obtain the two morphological forms rather than differences intrinsic to the cell types, we performed real-time RT-PCR experiments using RNAs derived from both yeast cells and mycelia that had been cultured at 37degreesC and 26degreesC in either complete medium (YPD or Sabouraud) or minimal medium. Twenty genes, including AGS1 (alpha-1,3-glucan synthase) and TSA1 (thiol-specific antioxidant), were shown to be more highly expressed in the yeast cells than in the hyphae. Although their levels of expression could be different in rich and minimal media, there was a general tendency for these genes to be more highly expressed in the yeast cells.
Palavra-chave Paracoccidioides brasiliensis
dimorphic fungi
suppression subtraction hybridization
macroarray analysis
real-time RT-PCR
Idioma Inglês
Data de publicação 2004-07-01
Publicado em Molecular Genetics and Genomics. New York: Springer, v. 271, n. 6, p. 667-677, 2004.
ISSN 1617-4615 (Sherpa/Romeo, fator de impacto)
Publicador Springer
Extensão 667-677
Fonte http://dx.doi.org/10.1007/s00438-004-1016-6
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000222677100004
Endereço permanente http://repositorio.unifesp.br/handle/11600/27815

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