Aviner Ranen, Lidsky Peter V, Xiao Yinghong, Tasseto Michel, Zhang Lichao, McAlpine Patrick L, Elias Joshua, Frydman Judith, Andino Raul
These authors contributed equally.
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94158, USA.
bioRxiv. 2023 Jul 6:2023.07.05.547902. doi: 10.1101/2023.07.05.547902.
A better mechanistic understanding of virus-host interactions can help reveal vulnerabilities and identify opportunities for therapeutic interventions. Of particular interest are essential interactions that enable production of viral proteins, as those could target an early step in the virus lifecycle. Here, we use subcellular proteomics, ribosome profiling analyses and reporter assays to detect changes in polysome composition and protein synthesis during SARS-CoV-2 (CoV2) infection. We identify specific translation factors and molecular chaperones whose inhibition impairs infectious particle production without major toxicity to the host. We find that CoV2 non-structural protein Nsp1 selectively enhances virus translation through functional interactions with initiation factor EIF1A. When EIF1A is depleted, more ribosomes initiate translation from an upstream CUG start codon, inhibiting translation of non-structural genes and reducing viral titers. Together, our work describes multiple dependencies of CoV2 on host biosynthetic networks and identifies druggable targets for potential antiviral development.
对病毒与宿主相互作用有更深入的机制理解，有助于揭示病毒的脆弱性，并为治疗干预找到机会。特别值得关注的是那些对病毒蛋白产生至关重要的相互作用，因为这些相互作用可能针对病毒生命周期的早期阶段。在此，我们利用亚细胞蛋白质组学、核糖体图谱分析和报告基因检测，来检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2，简称CoV2）感染过程中多核糖体组成和蛋白质合成的变化。我们鉴定出了特定的翻译因子和分子伴侣，抑制它们会损害感染性病毒颗粒的产生，且对宿主无重大毒性。我们发现，CoV2非结构蛋白Nsp1通过与起始因子EIF1A的功能性相互作用，选择性地增强病毒翻译。当EIF1A缺失时，更多核糖体从上游CUG起始密码子开始翻译，抑制非结构基因的翻译并降低病毒滴度。总之，我们的研究描述了CoV2对宿主生物合成网络的多种依赖性，并确定了潜在抗病毒药物开发的可成药靶点。
