Functional analysis of Paracoccidioides brasiliensis 14-3-3 adhesin expressed in Saccharomyces cerevisiae. 2015

Patricia Akemi Assato, and Julhiany de Fátima da Silva, and Haroldo Cesar de Oliveira, and Caroline Maria Marcos, and Danuza Rossi, and Sandro Roberto Valentini, and Maria José Soares Mendes-Giannini, and Cleslei Fernando Zanelli, and Ana Marisa Fusco-Almeida
Laboratório de Micologia Clínica - Núcleo de Proteômica - Faculdade de Ciências Farmacêuticas- Unesp - Campus Araraquara, Rodovia Araraquara - Jaú Km 1, 14801-902, Araraquara, SP, Brazil. patricia.assato@gmail.com.

BACKGROUND 14-3-3 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. RESULTS The 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. CONCLUSIONS Our 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.

UI MeSH Term Description Entries
D010228 Paracoccidioides A mitosporic fungal genus. P. brasiliensis (previously Blastomyces brasiliensis) is the etiologic agent of PARACOCCIDIOIDOMYCOSIS. Blastomyces brasiliensis,Loboa loboi,Paracoccidioides brasiliensis
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004875 Ergosterol A steroid occurring in FUNGI. Irradiation with ULTRAVIOLET RAYS results in formation of ERGOCALCIFEROL (vitamin D2). Lumisterol,Pro-Vitamin D2,Provitamin D 2,D2, Pro-Vitamin,Pro Vitamin D2
D005656 Fungal Proteins Proteins found in any species of fungus. Fungal Gene Products,Fungal Gene Proteins,Fungal Peptides,Gene Products, Fungal,Yeast Proteins,Gene Proteins, Fungal,Peptides, Fungal,Proteins, Fungal
D005816 Genetic Complementation Test A test used to determine whether or not complementation (compensation in the form of dominance) will occur in a cell with a given mutant phenotype when another mutant genome, encoding the same mutant phenotype, is introduced into that cell. Allelism Test,Cis Test,Cis-Trans Test,Complementation Test,Trans Test,Allelism Tests,Cis Tests,Cis Trans Test,Cis-Trans Tests,Complementation Test, Genetic,Complementation Tests,Complementation Tests, Genetic,Genetic Complementation Tests,Trans Tests
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D015966 Gene Expression Regulation, Fungal Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in fungi. Fungal Gene Expression Regulation,Regulation of Gene Expression, Fungal,Regulation, Gene Expression, Fungal
D048948 14-3-3 Proteins A large family of signal-transducing adaptor proteins present in wide variety of eukaryotes. They are PHOSPHOSERINE and PHOSPHOTHREONINE binding proteins involved in important cellular processes including SIGNAL TRANSDUCTION; CELL CYCLE control; APOPTOSIS; and cellular stress responses. 14-3-3 proteins function by interacting with other signal-transducing proteins and effecting changes in their enzymatic activity and subcellular localization. The name 14-3-3 derives from numerical designations used in the original fractionation patterns of the proteins. 14-3-3 Protein,14-3-3 Protein, beta Isoform,14-3-3 Protein, epsilon Isoform,14-3-3 Protein, eta Isoform,14-3-3 Protein, gamma Isoform,14-3-3 Protein, tau Isoform,14-3-3 Protein, zeta Isoform,14-3-3 beta Protein,14-3-3 epsilon Protein,14-3-3 eta Protein,14-3-3 gamma Protein,14-3-3 tau Protein,14-3-3 zeta Protein,14-3-3beta Protein,14-3-3epsilon Protein,14-3-3eta Protein,14-3-3gamma Protein,14-3-3tau Protein,14-3-3zeta Protein,Brain 14-3-3 Protein,Protein 14-3-3,14 3 3 Protein,14 3 3 Protein, beta Isoform,14 3 3 Protein, epsilon Isoform,14 3 3 Protein, eta Isoform,14 3 3 Protein, gamma Isoform,14 3 3 Protein, tau Isoform,14 3 3 Protein, zeta Isoform,14 3 3 Proteins,14 3 3 beta Protein,14 3 3 epsilon Protein,14 3 3 eta Protein,14 3 3 gamma Protein,14 3 3 tau Protein,14 3 3 zeta Protein,14 3 3beta Protein,14 3 3epsilon Protein,14 3 3eta Protein,14 3 3gamma Protein,14 3 3tau Protein,14 3 3zeta Protein,14-3-3 Protein, Brain,Brain 14 3 3 Protein,Protein, 14-3-3tau,eta Protein, 14-3-3
D020869 Gene Expression Profiling The determination of the pattern of genes expressed at the level of GENETIC TRANSCRIPTION, under specific circumstances or in a specific cell. Gene Expression Analysis,Gene Expression Pattern Analysis,Transcript Expression Analysis,Transcriptome Profiling,Transcriptomics,mRNA Differential Display,Gene Expression Monitoring,Transcriptome Analysis,Analyses, Gene Expression,Analyses, Transcript Expression,Analyses, Transcriptome,Analysis, Gene Expression,Analysis, Transcript Expression,Analysis, Transcriptome,Differential Display, mRNA,Differential Displays, mRNA,Expression Analyses, Gene,Expression Analysis, Gene,Gene Expression Analyses,Gene Expression Monitorings,Gene Expression Profilings,Monitoring, Gene Expression,Monitorings, Gene Expression,Profiling, Gene Expression,Profiling, Transcriptome,Profilings, Gene Expression,Profilings, Transcriptome,Transcript Expression Analyses,Transcriptome Analyses,Transcriptome Profilings,mRNA Differential Displays
D029701 Saccharomyces cerevisiae Proteins Proteins obtained from the species SACCHAROMYCES CEREVISIAE. The function of specific proteins from this organism are the subject of intense scientific interest and have been used to derive basic understanding of the functioning similar proteins in higher eukaryotes. Baker's Yeast Proteins,S cerevisiae Proteins

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