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Interaction between Human Respiratory Syncytial Virus Nucleoprotein and cellular protein MAGEA11

Grant number: 15/10241-0
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): August 01, 2015
Effective date (End): July 31, 2016
Field of knowledge:Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
Principal Investigator:Armando Morais Ventura
Grantee:Stephanie Santos Suehiro Arcos
Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Human respiratory syncytial virus (HRSV) belongs to the Paramyxoviridae family and has a negative sense single stranded RNA genome, with approximately 15,000 nucleotides that encode 11 proteins. The HRSV is one of the main causes of respiratory diseases in children and newborns in the world, but also affect the elderly and people with immune deficiency. Still there are no drugs or vaccines effective against HRSV. Thus, the search for therapeutic targets against HRSV is a priority, with the potential to characterize them studying the virus-cell interaction. In this sense, an experiment was carried out with yeast double-hybrid system, where one of the proteins, used as bait, was the viral nucleoprotein (N), evaluated against two human libraries. There was strong indication of interaction between N and the cellular protein MAGEA11, which acts as co-regulator of androgen receptors through its ligation to the motif FXXLF. We found this motif in the amino acid sequence of N, making possible that it recruits MAGEA11 to the apparatus associated with the viral genome transcription and replication. To test this hypothesis, we propose to verify if interaction occurs between N and MAGEA11 in mammalian cells, through the techniques of co-immunoprecipation and co-localization by immunofluorescence. If so, we will do a complementation test using the Split Gaussia luciferase system to demonstrate the closeness between N and MAGEA11 at the molecular level, and check if MAGEA11 expression inhibition by siRNA affects HRSV replication in cell cultures.

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