eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus

Viruses

Su Li¹  Shuo Feng¹  Jing-Han Wang¹  Wen-Rui He²  Hua-Yang Qin²  Hong Dong²  Lian-Feng Li²  Shao-Xiong Yu²  Yongfeng Li²  Hua-Ji Qiu¹ 

[1] State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China;

关键词: classical swine fever virus;    virus–host interactions;    NS5A;    eukaryotic elongation factor 1A;    internal ribosome entry site;   

DOI  :  10.3390/v7082833

来源: mdpi

【 摘 要 】

The NS5A protein of classical swine fever virus (CSFV) is involved in the RNA synthesis and viral replication. However, the NS5A-interacting cellular proteins engaged in the CSFV replication are poorly defined. Using yeast two-hybrid screen, the eukaryotic elongation factor 1A (eEF1A) was identified to be an NS5A-binding partner. The NS5A–eEF1A interaction was confirmed by coimmunoprecipitation, glutathione S-transferase (GST) pulldown and laser confocal microscopy assays. The domain I of eEF1A was shown to be critical for the NS5A–eEF1A interaction. Overexpression of eEF1A suppressed the CSFV growth markedly, and conversely, knockdown of eEF1A enhanced the CSFV replication significantly. Furthermore, eEF1A, as well as NS5A, was found to reduce the translation efficiency of the internal ribosome entry site (IRES) of CSFV in a dose-dependent manner, as demonstrated by luciferase reporter assay. Streptavidin pulldown assay revealed that eEF1A could bind to the CSFV IRES. Collectively, our results suggest that eEF1A interacts with NS5A and negatively regulates the growth of CSFV.

【 授权许可】

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