Interação entre a proteína celular hSlu7 e a proteína NS5 do vírus da febre amarela

• Autor(a) principal: Gomes, Arieli Fernanda Gavioli
• Data de Publicação: 2011
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Biblioteca Digital de Teses e Dissertações da FAMERP
• Texto Completo: http://bdtd.famerp.br/handle/tede/170
• topic: hSlu7; Vírus da Febre Amarela; Interação Proteína-proteína; NS5; Sistema Duplo-Híbrido; hSlu7; Protein-protein interaction; Cytoplasmic Sublocalization; NS5; Two-hybrid assays; CNPQ::CIENCIAS DA SAUDE

Introduction: The Yellow Fever is characterized by severe hepatitis, renal failure, hemorrhage, and rapid terminal events that lead to shock and death. This disease is caused by the infection with the Yellow Fever Virus (YFV), considered the prototype of the Flavivirus genus. Its mechanism of replication is not well known but includes interactions of viral RNA with cellular and viral proteins. The nonstructural protein 5 (NS5) is the largest and most conserved protein of the Flavivirus genus; it encodes RNA-dependent RNA polymerase (RdRp) domains, besides possessing many important functions during viral replication, such as genic regulation of host cells. The protein hSlu7 is a homologous human protein, which was isolated interacting with the U5 that is involved in the second the step of alternative splicing. The hSlu7 is a predominantly nuclear protein and participates at the alternative splicing, influencing the correct choice of the alternative AGs of exon 3' so that the spliceossome is able to bind and start alternative splicing. Objective: To characterize the interaction of the hSlu7 protein with the YFV-NS5 protein and its cellular localization during viral infection. Material and Method: We confirmed the interaction of various NS5 with human proteins by two-hybrid assays. Deletion mutants were constructed and co-transformed with hSlu7 in yeast to determine the minimal domain of NS5 required for interaction. The cellular localization of the hSlu7 fused with GFP during the response of vaccine strain 17D of YFV in cells Vero E6, marked with anti-NS4AB and anti-NS5 for detection of the infection was also tested. Results: hSlu7 interacts with initial and final portions of the RdRp and the cytoplasmic sublocalization of hSlu7 occurs in the cells infected with YFV. Conclusions: Our results suggest that hSlu7 interacts with the YFV by two-hybrid system and the cellular sublocalization occurs due to the presence of viral infection. Further studies using RNA interference should be addressed to confirm the cellular function of hSlu7 , and to evaluate which alterations that infected and uninfected cells will suffer with low levels of hSlu7.