title: Interaction of cellular poly(C)-binding protein 2 with nonstructural protein 1β is beneficial to Chinese highly pathogenic porcine reproductive and respiratory syndrome virus replication.
abstract: Non-structural protein1β (Nsp1β) of porcine reproductive and respiratory syndrome virus (PRRSV) has been recognized to be involved in suppressing the host innate immune response and mediating viral subgenomic mRNA transcription. In the present study, we have analyzed the interaction of Nsp1β of Chinese highly pathogenic PRRSV (HP-PRRSV) with cellular poly(C)-binding 2 (PCBP2) by means of the yeast two-hybrid screening in a pulmonary alveolar macrophages (PAMs) cDNA library and co-immunoprecipitation (Co-IP) assay. Our results indicated that the Nsp1β of the HP-PRRSV is able to bind and interact with cellular PCBP2 strongly in both the infected cells and plasmid transfected cells. Their minimal binding regions were identified to be the residues 85-203 aa (PCPβ and CTE domains) for the Nsp1β and the residues 96-168 aa (KH2 domain) for PCBP2, respectively. Next, we used confocal immunofluorescence analysis and discovered that, during PRRSV infection in MARC-145 cells and/or plasmid-transfected cells, the Nsp1β and PCBP2 mainly colocalized in the cytoplasm and perinuclear pattern. Moreover, the siRNA-mediated silencing of PCBP2 gene in the MARC-145 cells resulted in significant reduction of the virus titer in supernatants as well as viral proteins, while no significant effects on the expression of the type I interferon α and interferon β, suggesting that the interaction of the Nsp1β with cellular PCBP2 is beneficial to Chinese HP-PRRSV replication in MARC-145 cells.
authors.affiliation: Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.forename: Qing
authors.initials: Q
authors.lastname: He
authors.forename: Yueyi
authors.initials: Y
authors.lastname: Gao
authors.forename: Xin
authors.initials: X
authors.lastname: Guo
authors.forename: Xinna
authors.initials: X
authors.lastname: Ge
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.forename: Hanchun
authors.initials: H
authors.lastname: Yang
mesh_terms: D000818:Animals; D002460:Cell Line; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D008264:Macrophages / Q000821:virology; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D019316:Porcine respiratory and reproductive syndrome virus; D011485:Protein Binding; D025941:Protein Interaction Mapping; D016601:RNA-Binding Proteins / Q000378:metabolism*; D013552:Swine; D020798:Two-Hybrid System Techniques; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D016601:RNA-Binding Proteins; D017361:Viral Nonstructural Proteins