Caracterização do fator de transcrição AreA de P. lutzii
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Data
2024-11-29
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Universidade Federal de Goiás
Resumo
Paracoccidioidomycosis is a mycosis endemic to subtropical areas of Latin America, caused by
fungi of the genus Paracoccidioides. Infection occurs via inhalation of conidia or fragments of
mycelium. In the lungs of the host, phagocytic cells engage in the process of phagocytosis to
combat the microorganism. Within the phagosome, alterations in pH, oxidative stress, and
nutritional limitation present challenges to the pathogen's survival. In this context, nutritional
immunity can be defined as a process that deprives the pathogen of preferred sources of essential
nutrients required for the synthesis of biomolecules, such as nitrogen sources. Pathogenic fungi
such as Paracoccidioides spp. have a coordinated mechanism for capturing non-preferred sources
of nitrogen, which is known as Nitrogen Catabolic Repression (NCR). The transcription factor
GATA AreA is responsible for regulating this process, ensuring the catabolism of nitrogen available
in the environment by controlling the genes responsible for the uptake and assimilation of these
sources. The objective of this study was to examine the influence of areA gene silencing on the
assimilation of nitrogen sources, growth, cell wall composition, and cell size of P. lutzii, with
comparisons between mutant and wild-type strains grown in varying concentrations of glutamine.
The gap and areA genes were evaluated by RT-qPCR. The growth curve was measured by
absorbance at 0, 12, 24, and 48 hours. The viability of the cells was estimated by propidium iodide,
followed by a subsequent analysis under a fluorescence microscope. To evaluate the composition
of the cell wall and cell size, the cells were stained with calcofluor white and examined under a
fluorescence microscope. The gap and areA genes exhibited elevated expression levels in the
glutamine mutant strain. The CFW analysis indicates a discrepancy in cell wall composition
between the strains in the presence of 10 mM glutamine. Furthermore, the mutant exhibited
increased cell size under these conditions. Consequently, the results indicate that the strain in which
the areA gene has been silenced displays a distinct biological response when grown in 10 mM
glutamine, which differs from that described in the literature. An analysis of the expression of new
gene targets and an evaluation of the proteomic profile in the therefore mentioned conditions may
assist in identifying crucial factors for understanding the role of AreA in P. lutzii.
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Palavras-chave
Paracoccidioidomicose, Metabolismo de nitrogênio, Glutamina, AreA, Paracoccidioidomycosis, Nitrogen metabolism, Glutamine, AreA
Citação
PEIXOTO, M. E. D. Caracterização do fator de transcrição AreA de P. lutzii. 2024. 37 f. Trabalho de Conclusão de Curso (Bacharel em Biomedicina)- Instituto de Ciências Biológicas, Universidade Federal de Goiás, 2024.