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Eukaryotic Cell, February 2003, p. 34-48, Vol. 2, No. 1
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.1.34-48.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Expressed Sequence Tag Analysis of the Human Pathogen Paracoccidioides brasiliensis Yeast Phase: Identification of Putative Homologues of Candida albicans Virulence and Pathogenicity Genes

Gustavo H. Goldman,1* Everaldo dos Reis Marques,1 Diógenes Custódio Duarte Ribeiro,1 Luciano Ângelo de Souza Bernardes,1 Andréa Carla Quiapin,2 Patrícia Marostica Vitorelli,2 Marcela Savoldi,1 Camile P. Semighini,1 Regina C. de Oliveira,3 Luiz R. Nunes,3 Luiz R. Travassos,4 Rosana Puccia,4 Wagner L. Batista,4 Leslie Ecker Ferreira,5 Júlio C. Moreira,5 Ana Paula Bogossian,5 Fredj Tekaia,6 Marina Pasetto Nobrega,5 Francisco G. Nobrega,5 and Maria Helena S. Goldman2

Faculdade de Ciências Farmacêuticas de Ribeirão Preto,1 Faculdade de Filosofia, Ciências, e Letras de Ribeirão Preto, Universidade de São Paulo,2 Departamento de Microbiologia, Imunologia, e Parasitologia, Universidade Federal de São Paulo, São Paulo,4 Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes,3 Universidade do Vale do Paraíba, UNIVAP, Vale do Paraíba, Brazil,5 Unité de Génétique Moléculaire des Levures, Institut Pasteur, Paris, France6

Received 17 July 2002/ Accepted 23 October 2002

Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of the prevalent systemic mycosis in Latin America, paracoccidioidomycosis. We present here a survey of expressed genes in the yeast pathogenic phase of P. brasiliensis. We obtained 13,490 expressed sequence tags from both 5' and 3' ends. Clustering analysis yielded the partial sequences of 4,692 expressed genes that were functionally classified by similarity to known genes. We have identified several Candida albicans virulence and pathogenicity homologues in P. brasiliensis. Furthermore, we have analyzed the expression of some of these genes during the dimorphic yeast-mycelium-yeast transition by real-time quantitative reverse transcription-PCR. Clustering analysis of the mycelium-yeast transition revealed three groups: (i) RBT, hydrophobin, and isocitrate lyase; (ii) malate dehydrogenase, contigs Pb1067 and Pb1145, GPI, and alternative oxidase; and (iii) ubiquitin, delta-9-desaturase, HSP70, HSP82, and HSP104. The first two groups displayed high mRNA expression in the mycelial phase, whereas the third group showed higher mRNA expression in the yeast phase. Our results suggest the possible conservation of pathogenicity and virulence mechanisms among fungi, expand considerably gene identification in P. brasiliensis, and provide a broader basis for further progress in understanding its biological peculiarities.

* Corresponding author. Mailing address: 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. Phone: 0055-016-6024280/81. Fax: 0055-016-6331092. E-mail: ggoldman{at}usp.br.

Eukaryotic Cell, February 2003, p. 34-48, Vol. 2, No. 1
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.1.34-48.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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