title: Poly(A)-binding protein interacts with the nucleocapsid protein of porcine reproductive and respiratory syndrome virus and participates in viral replication.
abstract: Interactions between host factors and the viral protein play important roles in host adaptation and regulation of virus replication. Poly(A)-binding protein (PABP), a host cellular protein that enhances translational efficiency by circularizing mRNAs, was identified by yeast two-hybrid screening as a cellular partner for PRRSV nucleocapsid (N) protein in porcine alveolar macrophages. The specific interaction of PRRSV N protein with PABP was confirmed in infected cells by co-immunoprecipitation and in vitro by GST pull-down assay. We showed by confocal microscopy that the PABP co-localized with the PRRSV N protein. Using a series of deletion mutants, the interactive domain of N protein with PABP was mapped to a region of amino acids 52-69. For PABP, C-terminal half, which interestingly interacts other translation regulators, was determined to be the domain interactive with N protein. Short hairpin RNA (shRNA)-mediated silencing of PABP in cells resulted in significantly reduced PRRSV RNA synthesis, viral encoded protein expression and viral titer. Overall, the results presented here point toward an important role for PABP in regulating PRRSV replication.
authors.affiliation: Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.
authors.forename: Xiaoye
authors.initials: X
authors.lastname: Wang
authors.forename: Juan
authors.initials: J
authors.lastname: Bai
authors.forename: Lili
authors.initials: L
authors.lastname: Zhang
authors.forename: Xianwei
authors.initials: X
authors.lastname: Wang
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Li
authors.forename: Ping
authors.initials: P
authors.lastname: Jiang
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D020868:Gene Silencing; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D016676:Macrophages, Alveolar / Q000378:metabolism / Q000821:virology; D018613:Microscopy, Confocal; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D039102:Poly(A)-Binding Proteins / Q000378:metabolism*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D020798:Two-Hybrid System Techniques; D053585:Virus Attachment; D014779:Virus Replication
chemical_list: D019590:Nucleocapsid Proteins; D039102:Poly(A)-Binding Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral