Title:	Construção e caracterização de mutantes de cryptococcus neoformans com deleção de genes das vias biossíntéticas de fosfatidilcolina
Authors:	Timboni, Filipe dos Santos
Orientador(es)::	Matos, Larissa Fernandes
Assunto::	Cryptococcus neoformans Virulência (Microbiologia) Deleção gênica Vias biossintéticas Fosfatidilcolina
Issue Date:	12-Mar-2025
Data de defesa::	23-May-2024
Citation:	TIMBONI, Filipe dos Santos. Construção e caracterização de mutantes de cryptococcus neoformans com deleção de genes das vias biossíntéticas de fosfatidilcolina. 2024. 84 f. Dissertação (Mestrado em Biologia Microbiana) — Universidade de Brasília, Brasília, 2024.
Abstract:	Phospholipids are fundamental molecules for the structure and function of the plasma membrane in both eukaryotic and prokaryotic cells, as well as being involved in environmental adaptation processes and cell signaling. Currently, various studies explore phospholipid biosynthesis pathways as potential therapeutic targets against pathogenic fungi. Thus, this study aimed to investigate the role of phosphatidylcholine biosynthesis pathways in the virulence of the fungus Cryptococcus neoformans through the construction and characterization of knockou mutants for the OPI3 and PCT1 genes, which encode the enzymes of the CDP-DAG and Kennedy pathways, respectively. The mutants were obtained through molecular gene deletion techniques using Double Joint-Polymerase Chain Reaction (DJ-PCR) and biolistics, and were subjected to phenotypic tests to evaluate the main attributes associated with C. neoformans virulence. The mutant deleted for the OPI3 gene, from the de novo pathway, showed significantly affected growth in the absence of choline in the culture medium, while the double mutant for both pathways, opi3Δpct1Δ, was able to grow only in the presence of yeast extract or on egg yolk agar, possibly due to the assimilation of substrates used by a third pathway. Both opi3Δ and opi3Δpct1Δ mutants induced larger polysaccharide capsules than the wild-type KN99α, while in antifungal susceptibility tests, the pct1Δ mutant showed greater susceptibility to the azoles, fluconazole, and itraconazole. The results indicate that these pathways may be associated with cellular mechanisms that regulate polysaccharide capsule synthesis, as well as play a role in virulence and plasma membrane integrity.
metadata.dc.description.unidade:	Instituto de Ciências Biológicas (IB) Departamento de Biologia Celular (IB CEL)
metadata.dc.description.ppg:	Programa de Pós-Graduação em Biologia Microbiana
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Appears in Collections:	Teses, dissertações e produtos pós-doutorado

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