Classical swine fever (CSF) is a contagious disease with a high mortality rate and is caused by classical swine fever virus(CSFV). CSFV non-structural protein 4B (NS4B) plays a crucial role in CSFV replication and pathogenicity. However,precisely how NS4B exerts these functions remains unknown, especially as there are no reports relating to potential cellularpartners of CSFV NS4B. Here, a yeast two-hybrid (Y2H) system was used to screen the cellular proteins interacting withNS4B from a porcine alveolar macrophage (PAM) cDNA library. The protein screen along with alignment using the NCBIdatabase revealed 14 cellular proteins that interact with NS4B: DDX39B, COX7C, FTH1, MAVS, NR2F6, RPLP1,PSMC4, FGL2, MKRN1, RPL15, RPS3, RAB22A, TP53BP2 and TBK1. These proteins mostly relate to oxidoreductaseactivity, signal transduction, localization, biological regulation, catalytic activity, transport and metabolism by GO categories.Tank-binding kinase 1 (TBK1) was chosen for further confirmation. The NS4B-TBK1 interaction was furtherconfirmed by subcellular co-location, co-immunoprecipitation and glutathione S-transferase pull-down assays. This studyoffers a theoretical foundation for further understanding of the diversity of NS4B functions in relation to viral infection andsubsequent pathogenesis.
Volume 45, 2020
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