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Classical swine fever virus non-structural protein 4B binds tank-binding kinase 1

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Abstract

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 cellular partners of CSFV NS4B. Here, a yeast two-hybrid (Y2H) system was used to screen the cellular proteins interacting with NS4B from a porcine alveolar macrophage (PAM) cDNA library. The protein screen along with alignment using the NCBI database 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 oxidoreductase activity, 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 further confirmed by subcellular co-location, co-immunoprecipitation and glutathione S-transferase pull-down assays. This study offers a theoretical foundation for further understanding of the diversity of NS4B functions in relation to viral infection and subsequent pathogenesis.

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Acknowledgements

The research was supported by National Nature Science Foundation of China (No. 31472210) and Scientific Research Foundation of the Programs for Science and Technology Development of Henan Province, China (No. 162102110033).

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    Authors and Affiliations

    1. College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, China

      Huifang Lv, Wang Dong, Zhi Cao, Jihui Lin, Yueling Ouyang, Kangkang Guo & Yanming Zhang

    2. College of Pharmaceutical Engineering, Henan University of Animal Husbandry and Economy, No. 6 Longzihu Northern Road, Zhengzhou, 450046, Henan, China

      Huifang Lv & Cunfa Li

    3. College of Veterinary Medicine, Henan University of Animal Husbandry and Economy, No. 6 Longzihu Northern Road, Zhengzhou, 450046, Henan, China

      Wang Dong

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    1. Huifang Lv
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    Correspondence to Yanming Zhang.

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    Communicated by Saumitra Das.

    Corresponding editor: Saumitra Das

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    Lv, H., Dong, W., Cao, Z. et al. Classical swine fever virus non-structural protein 4B binds tank-binding kinase 1. J Biosci 43, 947–957 (2018). https://doi.org/10.1007/s12038-018-9802-1

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