Systems Biology Reveals NS4B-Cyclophilin A Interaction: A New Target to Inhibit YFV Replication

Autor(es)

Vidotto, Alessandra ; Morais, Ana T. S. ; Ribeiro, Milene R. ; Pacca, Carolina C. ; Terzian, Ana C. B. ; Gil, Laura H. V. G. ; Mohana-Borges, Ronaldo ; Gallay, Philippe ; Nogueira, Mauricio L.

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF PROTEOME RESEARCH; v. 16, n. 4, p. 1542-1555, APR 2017.
• Citações Web of Science: 6

Resumo

Yellow fever virus (YFV) replication is highly dependent on host cell factors. YFV NS4B is reported to be involved in viral replication and immune evasion. Here interactions between NS4B and human proteins were determined using a GST pull-down assay and analyzed using 1-DE and LC-MS/MS. We present a total of 207 proteins confirmed using Scaffold 3 Software. Cyclophilin A (CypA), a protein that has been shown to be necessary for the positive regulation of flavivirus replication, was identified as a possible NS4B partner. 59 proteins were found to be significantly increased when compared with a negative control, and CypA exhibited the greatest difference, with a 22-fold change. Fisher's exact test was significant for 58 proteins, and the p value of CypA was the most significant (0.000000019). The Ingenuity Systems software identified 16 pathways, and this analysis indicated sirolimus, an mTOR pathway inhibitor, as a potential inhibitor of CypA. Immunofluorescence and viral plaque assays showed a significant reduction in YFV replication using sirolimus and cyclosporine A (CsA) as inhibitors. Furthermore, YFV replication was strongly inhibited in treated with both inhibitors using reporter BHK-21-rep-YFV17DLucNeolres cells. Taken together, these data suggest that CypA-NS4B interaction regulates YFV replication. Finally, we present the first evidence that YFV inhibition may depend on NS4B-CypA interaction. (AU)

• Processo FAPESP: 09/01400-7 - ESTUDO DAS INTERAÇÕES ENTRE A PROTEÍNA NS4b DO VÍRUS DA FEBRE AMARELA E PROTEÍNAS CELULARES
• Beneficiário: Maurício Lacerda Nogueira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 04/14846-0 - Rede de proteoma do estado de São Paulo
• Beneficiário: Fabio Cesar Gozzo
• Modalidade de apoio: Auxílio à Pesquisa - Programa GENOMA
• Processo FAPESP: 10/05043-1 - Caracterização da interação entre proteínas não estruturais do vírus da Febre Amarela e eIF3L
• Beneficiário: Ana Theresa Silveira de Morais
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto