Abstract
Influenza A virus exploits multiple host proteins during infection. To define the virus–host interactome, our group conducted a proteomics-based screen and identified 299 genes that contributed to virus replication and 24 genes that were antiviral. Of these genes, we focused on the role during virus replication of eukaryotic elongation factor 1 gamma (eEF1G), which is a subunit of the eukaryotic elongation factor-1 complex and known to be a pro-viral host protein. Using the CRISPR/Cas9 system, we obtained two clones that were defective in eEF1G expression. In both of these clones, A/WSN/33 (H1N1) virus growth and protein expression were significantly suppressed, but viral mRNA, vRNA, and cRNA expression were not reduced. However, the replication and protein expression of A/California/04/2009 (H1N1pdm) virus in both clones were similar to those in parental cells. We found that the PB2 and PA proteins of WSN virus were responsible for the eEF1G-dependent replication. Our data show that eEF1G plays a role in the translation of virus proteins in a strain-specific manner. Additional analyses may be needed to further understand the role of strain-specific host proteins during virus replication.
Highlights
Influenza A virus possesses eight single-stranded, negative-sense RNAs as a genome (Palese and Schulman, 1976)
Since this defect in eukaryotic elongation factor-1 gamma (eEF1G) expression may affect its localization, we determined the intracellular localization of eEF1G in clones 1–24 and 3–9. eEF1G was distributed in the cytoplasm of wild-type A549 cells and clone 3–9, whereas eEF1G was weakly detected in the cytoplasm of clone 1–24 (Figure 1E)
Since eEF1G is a subunit of the eukaryotic elongation factor-1 complex, which is responsible for the enzymatic delivery of aminoacyl-tRNAs to the ribosome (Carvalho et al, 1984)
Summary
Influenza A virus possesses eight single-stranded, negative-sense RNAs as a genome (Palese and Schulman, 1976). To better understand the host–virus interaction during virus replication, eight genome-wide screens have been reported and each identified numerous host proteins that are required for virus replication (Brass et al, 2009; Shapira et al, 2009; Karlas et al, 2010; König et al, 2010; Ward et al, 2012; Su et al, 2013; Tran et al, 2013; Watanabe et al, 2014). Down-regulation of eEF1G expression reduced virus propagation in vitro, it was not clear which step of virus propagation eEF1G is involved in
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