title: Human respiratory syncytial virus N, P and M protein interactions in HEK-293T cells.
abstract: Characterization of Human Respiratory Syncytial Virus (HRSV) protein interactions with host cell components is crucial to devise antiviral strategies. Viral nucleoprotein, phosphoprotein and matrix protein genes were optimized for human codon usage and cloned into expression vectors. HEK-293T cells were transfected with these vectors, viral proteins were immunoprecipitated, and co-immunoprecipitated cellular proteins were identified through mass spectrometry. Cell proteins identified with higher confidence scores were probed in the immunoprecipitation using specific antibodies. The results indicate that nucleoprotein interacts with arginine methyl-transferase, methylosome protein and Hsp70. Phosphoprotein interacts with Hsp70 and tropomysin, and matrix with tropomysin and nucleophosmin. Additionally, we performed immunoprecipitation of these cellular proteins in cells infected with HRSV, followed by detection of co-immunoprecipitated viral proteins. The results indicate that these interactions also occur in the context of viral infection, and their potential contribution for a HRSV replication model is discussed.
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Andressa P
authors.initials: AP
authors.lastname: Oliveira
authors.forename: Fernando M
authors.initials: FM
authors.lastname: Simabuco
authors.forename: Rodrigo E
authors.initials: RE
authors.lastname: Tamura
authors.forename: Manuel C
authors.initials: MC
authors.lastname: Guerrero
authors.forename: Paulo G G
authors.initials: PG
authors.lastname: Ribeiro
authors.forename: Towia A
authors.initials: TA
authors.lastname: Libermann
authors.forename: Luiz F
authors.initials: LF
authors.lastname: Zerbini
authors.forename: Armando M
authors.initials: AM
authors.lastname: Ventura
mesh_terms: D057809:HEK293 Cells; D050884:HSP72 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D000090243:Nucleophosmin; D009698:Nucleoproteins / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism; D018357:Respiratory Syncytial Virus Infections / Q000378:metabolism* / Q000821:virology*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins
chemical_list: D050884:HSP72 Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D009698:Nucleoproteins; C529603:P protein, human respiratory syncytial virus; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; D015678:Viral Structural Proteins; D000090243:Nucleophosmin; C077013:PRMT2 protein, human; D011484:Protein-Arginine N-Methyltransferases
keywords: Human respiratory syncytial virus; Matrix; Nucleoprotein; Optimized genes; Phosphoprotein; Protein interaction