Análise proteômica da fase leveduriforme do fungo patogênico Paracoccidioides sp durante a privação de nitrogênio

• Ano de defesa: 2014
• Autor(a) principal: Cruz-Leite, Vanessa Rafaela Milhomem
• Orientador(a): Borges, Clayton Luiz
• Banca de defesa: Borges, Clayton Luiz, Bailão, Alexandre Melo, Brito, Wesley de Almeida, Casaletti, Luciana, Pereira, Maristela
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Goiás
• Programa de Pós-Graduação: Programa de Pós-graduação em Genetica e Biologia Molecular
• Departamento: Instituto de Ciências Biológicas - ICB (RG)
• País: Brasil
• Palavras-chave em Português: Paracoccidioides sp; Metabolismo de nitrogênio; Proteômica
• Palavras-chave em Inglês: Proteomics; Nitrogen metabolism
• Área do conhecimento CNPq: GENETICA::GENETICA MOLECULAR E DE MICROORGANISMOS
• Link de acesso: http://repositorio.bc.ufg.br/tede/handle/tede/4543

The fungi of the genus Paracoccidioides sp are etiologic agents of disease paracoccidioidomycosis (PCM), are termodimórficos and have the ability to move the saprobiótica form to yeast form at a temperature around 37 ° C. The uptake is essential for the growth and adaptation of the fungus in host tissue, where nitrogen-dependent pathways have close relationship with pathogenicity. Pathogenic organisms possess a regulatory system called Nitrogen Catabolic Represion which is induced when nitrogen availability is limiting in surround causing the expression of genes necessary for nutrient uptake when preferred sources such as glutamine and ammonia are scarce. This regulatory process comprises a complex system in the infectious process. This study aims to identify proteins regulated by nitrogen depletion, where the pathogenic fungus Pb01-like was analyzed as a proteomic response. Pb01-like was grown in minimal medium control MMcM (+N) and treated MMcM (-N) for 6 hours at 37 ° C, cytoplasmic proteins were extracted and subjected to tryptic digestion and quantification. The proteomic profile revealed an expression of 135 proteins, 40 proteins induced or identified the treaty, 58 repressed or identified control and 44 proteins were expressed constitutively. In silico analyzes showed that the gene is regulated by formamidase areA transcription factor that responds conditions of nitrogen depletion in Aspergillus nidulans and the surrounding medium the enzymatic activity for the gene of formamidase Pb01-like induced depletion of nitrogen during. The amino acid sequence of the regulatory gene areA important in the metabolism of nitrogen was aligned between Pb01-like, Pb18, Aspergillus nidulans and Neurospora crasssa where this alignment showed high identity and homology of conserved zinc finger domains and DUF 1752. Metabolic pathways such as fermentation, gluconeogenesis, protein synthesis, nitrogen metabolism were induced for the depletion of nitrogen, showing a possible modulation of the metabolism of the fungus in order to be able to adapt to environments with nitrogen limitation.