Effect of N-glycosylation inhibition on the transcriptional and proteomic profiles of Paracoccidioides brasiliensis

• Grant number: 13/10741-8
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2013
• End date: June 30, 2016
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Maria Cristina Roque Antunes Barreira
• Grantee: Fausto Bruno dos Reis Almeida
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 13/26382-7 - Role of galectin-3 on the extracellular vesicles secretion from Cryptococcus neoformans, BE.EP.PD

Abstract

Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, the most prevalent systemic mycosis in Latin America. Fungus can able to increase their pathogenicity by factors as lipids, polysaccharides and glycoproteins. Glycosylation is important for protein secretion, function, stability and immunogenicity. N-glycans linked to proteins have been characterized in many species of yeast to mammals. In P. brasiliensis yeast, N-glycans play important role in the activity of N-acetyl-beta-D-glucosaminidase (NAGase) produced by yeast, and are directly involved in fungal growth and morphogenesis. N-glycans also exert considerable influence on the regulation of expression of NAGase, ²-1,3-glucanase, ±-1,4-amylase and other enzymes related to biosynthesis/degradation of the P. brasiliensis cell wall. Due the fact of N-glycans influence on the biological activities of P. brasiliensis proteins, have motivated us to investigate the effects of inhibition of N-glycosylation on the transcriptome and proteome of yeast. Therefore, we propose to do the following aims: (1) evaluate the effect of inhibition of N-glycosylation (provided by using tunicamycin) on the transcriptional profile of P. brasiliensis by HSS (Subtractive Hybridization deleted), (2) validate the differentially expressed genes by real-time PCR (RT-qPCR), (3) evaluate the inhibition effect of N-glycosylation on the proteomic profile of P. brasiliensis by 2D-DIGE (two-dimensional - Differential Gel Electrophoresis). Our expectation is that the achievement of these aims will allow the elucidation of the existence of a direct association between the N-glycosylation and regulation of genes that coding to proteins involved on growth, morphogenesis and pathogenicity of P. brasiliensis. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ALMEIDA, FAUSTO; SARDINHA-SILVA, ALINE; DA SILVA, THIAGO APARECIDO; PESSONI, ANDRE MOREIRA; PINZAN, CAMILA FIGUEIREDO; PAIVA ALEGRE-MALLER, ANA CLAUDIA; CECLIO, NERRY TATIANA; MORETTI, NILMAR SILVIO; LIMA DAMSIO, ANDRE RICARDO; PEDERSOLI, WELLINGTON RAMOS; et al. Toxoplasma gondii Chitinase Induces Macrophage Activation. PLoS One, v. 10, n. 12, DEC 10 2015. (13/10741-8)

ALMEIDA, FAUSTO; CAMPOS ANTONIETO, AMANDA CRISTINA; PESSONI, ANDRE MOREIRA; MONTEIRO, VALDIRENE NEVES; PAIVA ALEGRE-MALLER, ANA CLAUDIA; PIGOSSO, LAURINE LACERDA; PEREIRA, MARISTELA; DE ALMEIDA SOARES, CELIA MARIA; ROQUE-BARREIRA, MARIA CRISTINA. Influence of N-glycans on Expression of Cell Wall Remodeling Related Genes in Paracoccidioides brasiliensis Yeast Cells. CURRENT GENOMICS, v. 17, n. 2, p. 112-118, 2016. (13/10741-8)

DA SILVA, THIAGO APARECIDO; ROQUE-BARREIRA, MARIA CRISTINA; CASADEVALL, ARTURO; ALMEIDA, FAUSTO. Extracellular vesicles from Paracoccidioides brasiliensis induced M1 polarization in vitro. SCIENTIFIC REPORTS, v. 6, OCT 24 2016. (16/03322-7, 13/10741-8, 13/04088-0, 16/04877-2)

FREITAS, MATEUS S.; OLIVEIRA, ALINE F.; DA SILVA, THIAGO A.; FERNANDES, FABRICIO F.; GONCALES, RELBER A.; ALMEIDA, FAUSTO; ROQUE-BARREIRA, MARIA C.. Paracoccin Induces M1 Polarization of Macrophages via Interaction with TLR4. FRONTIERS IN MICROBIOLOGY, v. 7, JUL 5 2016. (13/10741-8, 12/09611-0)

OLIVEIRA, ALINE FERREIRA; FERNANDES, FABRICIO FREITAS; MARIANO, VANIA SAMMARTINO; ALMEIDA, FAUSTO; RUAS, LUCIANA PEREIRA; OLIVEIRA, LEANDRO LICURSI; OLIVER, CONSTANCE; JAMUR, MARIA CELIA; ROQUE-BARREIRA, MARIA CRISTINA. Paracoccin distribution supports its role in Paracoccidioides brasiliensis growth and dimorphic transformation. PLoS One, v. 12, n. 8, AUG 28 2017. (13/10741-8, 06/60642-2, 09/51197-3)