Functional analysis of Paracoccidioides brasiliensis 14-3-3 adhesin expressed in Saccharomyces cerevisiaeReport as inadecuate




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BMC Microbiology

, 15:256

Microbe-host interactions and microbial pathogenicity

Abstract

Background14-3-3 proteins comprise a family of eukaryotic multifunctional proteins involved in several cellular processes. The Pb14-3-3 of Paracoccidioides brasiliensis seems to play an important role in the Paracoccidioides-host interaction. Paracoccidioides brasiliensis is an etiological agent of paracoccidioidomycosis, which is a systemic mycosis that is endemic in Latin America. In the initial steps of the infection, Paracoccidioides spp. synthetizes adhesins that allow it to adhere and invade host cells. Therefore, the aim of this work was to perform a functional analysis of Pb14-3-3 using Saccharomyces cerevisiae as a model.

ResultsThe functional analysis of Pb14-3-3 was performed in S. cerevisiae, and it was found that Pb14-3-3 partially complemented S. cerevisiae proteins Bmh1p and Bmh2p, which are recognized as two yeast 14-3-3 homologues. When we evaluated the adhesion profile of S. cerevisiae transformants, Pb14-3-3 acted as an adhesin in S. cerevisiae; however, Bmh1p did not show this function. The influence of Pb14-3-3 in S. cerevisiae ergosterol pathway was also evaluated and our results showed that Pb14-3-3 up-regulates genes involved in ergosterol biosynthesis.

ConclusionsOur data showed that Pb14-3-3 was able to partially complement Bmh1p and Bmh2p proteins in S. cerevisiae; however, we suggest that Pb14-3-3 has a differential role as an adhesin. In addition, Pb-14-3-3 may be involved in Paracoccidioides spp. ergosterol biosynthesis which makes it an interest as a therapeutic target.

KeywordsParacoccidioides brasiliensis 14-3-3 protein Adhesion Adhesin AbbreviationsACOaconitase

ATCCAmerican Type Culture Collection

ECMextracellular matrix

ENOenolase

FZfluconazole

GAPDHglyceraldehyde-3-phosphate-dehydrogenase

Gp4343 kDa glycoprotein

ICLisocitrate lyase

LBLuria-Bertani

MLSmalate synthase

Pb14-3-314-3-3 protein from Paracoccidioides brasiliensis

PCRpolymerase chain reaction

SD-URAsynthetic defined medium without uracil

TPItriose phosphate isomerase

wtwild type

YEPDyeast extract peptone dextrose

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Author: Patricia Akemi Assato - Julhiany de Fátima da Silva - Haroldo Cesar de Oliveira - Caroline Maria Marcos - Danuza Rossi -

Source: https://link.springer.com/







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