Caracterização de anidrases carbônicas do fungo Paracoccidioides brasiliensis
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2011-08-29
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Universidade Federal de Goiás
Resumo
Paracoccidioidomycosis (PCM) is the most important endemic deep mycosis in Latin
America. Understanding of the complex interactions between the fungus and host must
include the identification of gene expression patterns during infection. Carbonic anhydrase
(CA) belongs to the family of zinc metalloenzymes that catalyzes the reversible hydratation of
carbon dioxide to bicarbonate. Transcriptional studies have shown that carbonic anhydrase of
P. brasiliensis is expressed in yeast cells recovered from liver of infected mice. In the present
work, we characterized the cDNAs encoding for four carbonic anhydrases of P.brasiliensis
(PbCA1, PbCA2, PbCA3, PbCA4). Recombinant PbCA1, 3 and 4 were obtained in
heterologous systems with 33 kDa, 28 kDa and 32 kDa respectively. Mass spectrometry
analysis confirmed the sequences of the produced proteins. The expression of PbCAs
transcripts was evaluated by using real-time RT-PCR in mycelium, yeast cells and mycelium
to yeast transition, in yeast cells exposed to CO2 and yeast cells recovery directly from liver
and spleen. In the presence of CO2, PbCA1, PbCA2 and PbCA4 gene expression was reduced
in the course of time. PbCA1 transcript expression was induced during the mycelium to yeast
transition. PbCA2 and PbCA4 gene expression was higher in yeast cells, when compared to
mycelium and mycelium to yeast transition. Pbca1 was induced yeast cells recovery directly
from liver and spleen while the transcripts for Pbca2 and Pbca4 were repressed in all
condictions. The gene expression data suggest differential roles of the CAs in the fungal
physiology.
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TOMAZETT, M. V. Caracterização de anidrases carbônicas do fungo Paracoccidioides brasiliensis. 2011. 47 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2011.