XI International Meeting on Paracoccidioidomycosis

XI International Meeting on Paracoccidioidomycosis

Resume:69-2

Oral / Poster

69-2	Trancriptional analysis of Paracoccidioides brasiliensis after exposition to itraconazole
Authors:	Benedito Rodrigues da Silva Neto (UFG - Universidade Federal de Goiás) ; Patricia Fernanda Zambuzzi Carvalho (UFG - Universidade Federal de Goiás) ; Nayche Santiago Pacheco de Santana (UFG - Universidade Federal de Goiás) ; Celia Maria de Almeida Soares (UFG - Universidade Federal de Goiás) ; Maristela Pereira (UFG - Universidade Federal de Goiás)

Abstract

Paracoccidioidomycosis (PCM) is a human illness endemic from Latin America. This disease is characterized by a granulomatous inflammation with clinical forms ranging from a benign localized infection to a disseminated one, affecting several parts of the body. About 10 million people are infected by the agent cause of this disease, a thermally dimorphic fungal pathogen, Paracoccidioides brasiliensis. The disease is acquired through of the inhalation of fungal propagules that reach the lungs where occurs the conversion from mycelial to the yeast phase. Itraconazole is a triazole antifungal drug, which is multiringed synthetic compounds containing three nitrogen atoms in the azole ring. The triazole drugs are broad-spectrum antifungal agents and are currently used to treat infections caused by various pathogenic yeast and molds. The mechanism of action of azoles has been elucidated to some fungi. They act by blocking the ergosterol, an essential cell membrane component, biosynthetic pathway through binding to and inhibition of the lanosterol 14-&Alphaα demethylase enzyme, encoded by the erg11. Here we aim to investigate the mechanism of action of itraconazole on P. brasiliensis, by using Representational Difference Analysis (RDA). P. brasiliensis yeast cells were grown on minimum medium in the presence and absence of itraconazole for 1 and 2 hour. ESTs sequences were clustering using the CAP3 program. By using the Blast2GO program ESTs were classified in agreement with the possible functions. The database sequence matches were considered significant at E-values ≤10-10. The analyses indicated the classification of transcripts in different functional category. Most of the regulated genes were involved in lipid metabolism including the precursors of ergosterol. The genes ERG11, ERG6, ERG3 and ERG5 were up regulated. In addition, were found genes involved in cell stress response, drug efflux, small molecule transport, elongation and transcription factors, cell wall and membrane, and hypothetic proteins. The up and down regulated genes by RDA and bioinformatics analysis were confirmed by real-time PCR.਀䘀椀渀愀渀挀椀愀氀匀甀瀀瀀漀爀琀㨀䌀一倀焀Ⰰ 䘀䤀一䔀倀攀䤀䘀匀⸀

Keyword: Paracoccidioides brasiliensis, Itraconazole, Transcriptional profile

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