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

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Abstract

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 37°C and 26°C in either complete medium (YPD or Sabouraud) or minimal medium. Twenty genes, including AGS1 (α-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.

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Acknowledgements

We would like to thank the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and MCT-PRONEX, Brazil, for financial support. We also thank the two anonymous reviewers for useful comments

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Authors and Affiliations

  1. Departamento de Ciências Farmacêuticas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café S/N, CEP 14040-903, Ribeirão Preto, São Paulo, Brazil

    E.R. Marques, M. E. S. Ferreira, M. Savoldi & G. H. Goldman

  2. Departamento de Genética e Evolução, Instituto de Biología, Universidade Estadual de Campinas (UNICAMP), CEP 13084-971, Campinas, São Paulo, Brazil

    R. D. Drummond, J. M. Felix & M. Menossi

  3. Departamento de Microbiología, Imunología e Parasitología, Universidade Federal de São Paulo, CEP-04023-062, São Paulo, Brazil

    L. R. Travassos, R. Puccia, W. L. Batista & K. C. Carvalho

  4. Departamento de Biología, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, CEP 14040-901, Ribeirão Preto, Brazil

    M. H. S. Goldman

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  1. E.R. Marques
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  2. M. E. S. Ferreira
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  3. R. D. Drummond
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  4. J. M. Felix
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  6. M. Savoldi
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  7. L. R. Travassos
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  9. W. L. Batista
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  10. K. C. Carvalho
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  11. M. H. S. Goldman
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  12. G. H. Goldman
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Correspondence to G. H. Goldman.

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Communicated by C P. Hollenberg

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Marques, E., Ferreira, M.E.S., Drummond, R.D. et al. Identification of genes preferentially expressed in the pathogenic yeast phase of Paracoccidioides brasiliensis, using suppression subtraction hybridization and differential macroarray analysis. Mol Genet Genomics 271, 667–677 (2004). https://doi.org/10.1007/s00438-004-1016-6

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