eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus.
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.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China.
Su
S
Li
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. fengshuoqa@163.com.
Shuo
S
Feng
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. lanyunfei901208@sina.cn.
Jing-Han
JH
Wang
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. wrhe0111@163.com.
Wen-Rui
WR
He
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. ziqi317@163.com.
Hua-Yang
HY
Qin
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. dong.hong55555@163.com.
Hong
H
Dong
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. lilianfeng124603@163.com.
Lian-Feng
LF
Li
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. yushaoxiong1987@hotmail.com.
Shao-Xiong
SX
Yu
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. yfli@hvri.ac.cn.
Yongfeng
Y
Li
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, China. huajiqiu@hvri.ac.cn.
Hua-Ji
HJ
Qiu
D000818:Animals; D002460:Cell Line; D002498:Centrifugation; D006692:Classical Swine Fever Virus / Q000276:immunology* / Q000502:physiology*; D015977:Eukaryotic Initiation Factor-1 / Q000378:metabolism*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D011485:Protein Binding; D025941:Protein Interaction Mapping; D013552:Swine; D020798:Two-Hybrid System Techniques; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication
D015977:Eukaryotic Initiation Factor-1; C096041:NS5 protein, flavivirus; D017361:Viral Nonstructural Proteins
NS5A; classical swine fever virus; eukaryotic elongation factor 1A; internal ribosome entry site; virus–host interactions