Pigs are an important host of influenza A viruses due to their ability to generate reassortant viruses with pandemic potential. NS1
protein of influenza A viruses is a key
virulence factor and a major antagonist of innate immune responses. It is also involved in enhancing viral mRNA translation and regulation of virus replication. Being a
protein with pleiotropic functions, NS1 has a variety of cellular interaction partners. Hence, studies on
swine influenza viruses (SIV) and identification of
swine influenza NS1-interacting host
proteins is of great interest. Here, we constructed a recombinant SIV carrying a
Strep-tag in the NS1
protein and infected primary swine respiratory epithelial cells (SRECs) with this virus. The
Strep-tag sequence in the NS1
protein enabled us to purify intact, the NS1
protein and its interacting
protein complex specifically. We identified cellular
proteins present in the purified complex by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and generated a dataset of these
proteins. 445
proteins were identified by LC-MS/MS and among them 192
proteins were selected by setting up a threshold based on MS parameters. The selected
proteins were analyzed by bioinformatics and were categorized as belonging to different functional groups including translation, RNA processing, cytoskeleton, innate immunity, and apoptosis. Protein interaction networks were derived using these data and the NS1 interactions with some of the specific host factors were verified by immunoprecipitation. The novel
proteins and the networks revealed in our study will be the potential candidates for targeted study of the molecular interaction of NS1 with host
proteins, which will provide insights into the identification of new therapeutic targets to control
influenza infection and disease pathogenesis.