Made available in DSpace on 2014-07-29T15:25:12Z (GMT). No. of bitstreams: 1 patricia kott.pdf: 7658177 bytes, checksum: ad9ad7831fb8596c7e55d3b1f0eee117 (MD5) Previous issue date: 2010-02-26 / Paracoccidioides brasiliensis is a thermo-dimorphic human pathogenic fungus that lives at 23”C in the mycelium phase (infecting phase) and at 37”C in the yeast phase (parasite phase). In attempt to survive, the cell wall of fungi can change its composition and/or structure in response to environmental stress by compensatory mechanisms. The molecules involved in these mechanisms are possible target for the development of effective antifungal agents. In P. brasiliensis, the main components of the cell wall are glucans and chitin polymers. These polymers make a primary barrier that is responsible for the structural integrity and form of the cell wall. In this work the behavior of P. brasiliensis was evaluated against stress conditions with the aim of study, for the first time, the mechanisms used by this fungus in the maintenance of the cell wall integrity. Our results shown that P. brasiliensis yeast cells are sensitive to cell wall stressors calcofluor white (CFW), congo red (CR), SDS, KCl, NaCl and sorbitol. There was an increase in the PbFKS1 transcripts expression and in the content of cell wall β- 1,3-glicana after treatment with all stressor agents. After treatment with SDS and KCl the PbGFA1 transcripts expression and the cell wall GlcNAc residues also increased. The transcript expression of PbGEL3 was also evaluated being increased after treatment with CFW, NaCl and sorbitol. Thus we showed that these molecules are involved in the maintenance of the cell wall against stress conditions. Apart from these analyses we obtained the active recombinant protein PbFks1pc. Through the anti-PbFks1pc antibody we performed immunocitolocalization assays. These experiments revealed the localization of PbFks1p in regions of apical growth in the mycelium phase and in the cell surface in yeast phase. / Paracoccidioides brasiliensis Š um fungo patog‰nico e termodim‚rfico que se apresenta na forma de micŠlio a 23”C (forma infectante) e na forma de levedura a 37”C (forma parasit€ria). A parede celular de fungos Š capaz de alterar sua composiƒ o e atŠ mesmo sua estrutura em resposta a condiƒ•es ambientais de estresse atravŠs de mecanismos compensat‚rios. As prote…nas envolvidas nestes mecanismos s o poss…veis alvos para o desenvolvimento de agentes antifˆngicos espec…ficos. Em P. brasiliensis, os principais pol…meros que constituem a parede celular s o glicanas e quitina. Estes pol…meros formam uma barreira prim€ria que Š respons€vel pela integridade estrutural e forma da parede celular. Neste trabalho, o comportamento de P. brasiliensis diante de situaƒ•es de estresse foi investigado com o intuito de estudar, pela primeira vez, os mecanismos utilizados por este fungo na manutenƒ o da integridade da parede celular. Nossos resultados mostram que cŠlulas leveduriformes de P. brasiliensis s o sens…veis aos agentes estressores da parede celular calcofluor white (CFW), congo red (CR), SDS, NaCl, KCl e sorbitol. Houve um aumento na express o de transcritos de PbFKS1 e no conteˆdo do pol…mero de β-1,3-glicana na parede celular ap‚s tratamento com todos os agentes estressores. Ap‚s tratamento com SDS e KCl a express o de transcritos de PbGFA1 assim como o conteˆdo de res…duos de GlcNAc na parede celular tambŠm aumentaram. A express o dos transcritos de PbGEL3 tambŠm foi avaliada estando aumentada ap‚s tratamento com CFW, NaCl e sorbitol. Desta forma, mostramos que estas molŠculas est o envolvidas na manutenƒ o da parede celular diante de situaƒ•es de estresse. AlŠm destas an€lises, a prote…na recombinante PbFks1pc ativa foi obtida. AtravŠs do anticorpo anti-PbFks1pc realizamos ensaios de imunocitolocalizaƒ o. Estes experimentos revelaram a presenƒa de PbFks1p em regi•es de crescimento apical no micŠlio e na superf…cie celular em cŠlulas leveduriformes.