The eukaryotic translation initiation factor 3 subunit E binds to classical swine fever virus NS5A and facilitates viral replication

Classical swine fever virus (CSFV) NS5A protein is a multifunctional protein, playing critical roles in viral RNA replication, translation and assembly. To further explore its functions in viral replication, interaction of NS5A with host factors was assayed using a his-tag “pull down” assay coupled...

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Bibliographic Details
Published in:Virology (New York, N.Y.) N.Y.), 2018-02, Vol.515, p.11-20
Main Authors: Liu, Xiaofeng, Wang, Xiaoyu, Wang, Qian, Luo, Mingyang, Guo, Huancheng, Gong, Wenjie, Tu, Changchun, Sun, Jinfu
Format: Article
Language:English
Subjects:
Animals
Cell Line
Classical Swine Fever - virology
Classical swine fever virus - genetics
Classical swine fever virus - physiology
CSFV
Down-Regulation
eIF3E
Eukaryotic Initiation Factor-3 - genetics
Eukaryotic Initiation Factor-3 - metabolism
Gene Expression
Genes, Reporter
Host-Pathogen Interactions
Interaction
NS5A
Replication
RNA, Small Interfering
RNA, Viral - genetics
Swine
Viral Nonstructural Proteins - genetics
Viral Nonstructural Proteins - metabolism
Virus Replication
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Summary:Classical swine fever virus (CSFV) NS5A protein is a multifunctional protein, playing critical roles in viral RNA replication, translation and assembly. To further explore its functions in viral replication, interaction of NS5A with host factors was assayed using a his-tag “pull down” assay coupled with shotgun LC-MS/MS. Host protein translation initiation factor 3 subunit E was identified as a binding partner of NS5A, and confirmed by co-immunoprecipitation and co-localization analysis. Overexpression of eIF3E markedly enhanced CSFV genomic replication, viral protein expression and production of progeny virus, and downregulation of eIF3E by siRNA significantly decreased viral proliferation in PK-15 cells. Luciferase reporter assay showed an enhancement of translational activity of the internal ribosome entry site of CSFV by eIF3E and a decrease in cellular translation by NS5A. These data indicate that eIF3E plays an important role in CSFV replication, thereby identifying it as a potential target for inhibition of the virus. [Display omitted] •CSFV NS5A protein interacts with host protein translation initiation factor 3 subunit E (eIF3E).•eIF3E co-localizes with CSFV NS5A and viral dsRNA in cells.•eIF3E positively modulates CSFV replication.•eIF3E enhance CSFV IRES translational efficiency.•NS5A inhibit host translation.
ISSN:0042-6822
1096-0341
DOI:10.1016/j.virol.2017.11.019