Interactome Profile of the Host Cellular Proteins and the Nonstructural Protein 2 of Porcine Reproductive and Respiratory Syndrome Virus

Abstract

The nonstructural protein 2 (NSP2) is considered to be one of crucial viral proteins in the replication and pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV). In the present study, the host cellular proteins that interact with the NSP2 of PRRSV were immunoprecipitated with anti-Myc antibody from the MARC-145 cells infected by a recombinant PRRSV with 3xMyc tag insertion in its NSP2-coding region, and then 285 cellular proteins interacting with NSP2 were identified by LC-MS/MS. The Gene Ontology and enriched KEGG Pathway bioinformatics analyses indicated that the identified proteins could be assigned to different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with infectious disease, translation, immune system, nervous system and signal transduction. Two interested cellular proteins–BCL2-associated athanogene 6 (BAG6) and apoptosis-inducing factor 1 (AIF1) which may involve in transporting of NSP2 to Endoplasmic reticulum (ER) or PRRSV-driven apoptosis were validated by Western blot. The interactome data between PRRSV NSP2 and cellular proteins contribute to the understanding of the roles of NSP2 in the replication and pathogenesis of PRRSV, and also provide novel cellular target proteins for elucidating the associated molecular mechanisms of the interaction of host cellular proteins with viral proteins in regulating the viral replication.

Porcine reproductive and respiratory syndrome virus nonstructural protein 2 (nsp2) topology and selective isoform integration in artificial membranes

“…The physical separation of the large cytoplasmic PLP2 protease domain and the luminal C-termini by the membrane interface would require that polyprotein processing of the nsp2 ↓ nsp3 junction to be cleaved either in trans following membrane insertion, or that cleavage of nsp2 ↓ nsp3 precedes membrane insertion. Similarly, current data supports both as possibilities; that the PLP2 exhibits both cis-and trans-cleavage activities (Han et al, 2009) and that nsp2 has been also shown to interact with cellular chaperones known to participate in post-translational membrane integration (Wang et al, 2014). Most simply, any nsp2 isoform resulting from a cleavage event between the PLP2 protease domain and the TM region would allow for physical separation of these domains and thus differential localization.…”

“…Since strong viral proteinÀ protein interactions can complicate assessment of native transmembrane insertion (van der Meer et al, 1998), and additionally PRRSV nsp2 interacts with a wide range of cellular proteins (Wang et al, 2014), we chose to complete these initial studies on nsp2 transmembrane functionality within a reductionist cell-free translation system in the absence of all other viral proteins.…”

Evaluation of bovine viral diarrhea virus interactions with its host proteome during the course of infection

“…As a result, we see few binding host proteins and pathways identified later after this stage that emphasize the CP-induced cytopathic effect. In addition, cellular proteins and pathways, such as protein binding, translation, and apoptosis, targeted by CP NS3 are known to be targeted by other viruses, indicating common strategies among these viruses [17,35].…”

Quantitative interactome reveals that porcine reproductive and respiratory syndrome virus nonstructural protein 2 forms a complex with viral nucleocapsid protein and cellular vimentin

“…Wang et al recently employed an IP approach combined with the LC-MS/MS to identify host cellular proteins that interact with PRRSV nsp2 [47]. Altogether, they identified 285 cellular proteins and selected two proteins to confirm these interactions.…”