Sammaibashi Shuhei, Yamayoshi Seiya, Kawaoka Yoshihiro
Division of Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, United States.
Front Microbiol. 2018 Jun 29;9:1446. doi: 10.3389/fmicb.2018.01446. eCollection 2018.
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.
甲型流感病毒在感染过程中利用多种宿主蛋白。为了确定病毒-宿主相互作用组，我们团队进行了一项基于蛋白质组学的筛选，鉴定出299个有助于病毒复制的基因和24个具有抗病毒作用的基因。在这些基因中，我们重点研究了真核延伸因子1γ（eEF1G）在病毒复制过程中的作用，它是真核延伸因子-1复合体的一个亚基，已知是一种促进病毒感染的宿主蛋白。利用CRISPR/Cas9系统，我们获得了两个eEF1G表达有缺陷的克隆。在这两个克隆中，A/WSN/33（H1N1）病毒的生长和蛋白表达均受到显著抑制，但病毒mRNA、vRNA和cRNA的表达并未降低。然而，A/California/04/2009（H1N1pdm）病毒在这两个克隆中的复制和蛋白表达与亲代细胞中的情况相似。我们发现WSN病毒的PB2和PA蛋白负责eEF1G依赖性复制。我们的数据表明，eEF1G以毒株特异性的方式在病毒蛋白的翻译过程中发挥作用。可能需要进一步的分析来深入了解毒株特异性宿主蛋白在病毒复制过程中的作用。
