Department of Microbiology and Immunology, Stanford University, Stanford, CA, USA.
INSERM U1110, Institute of Viral and Liver Diseases, University of Strasbourg, Strasbourg, France.
Nat Microbiol. 2019 Dec;4(12):2369-2382. doi: 10.1038/s41564-019-0518-2. Epub 2019 Aug 5.
Flaviviruses, including dengue virus (DENV) and Zika virus (ZIKV), cause severe human disease. Co-opting cellular factors for viral translation and viral genome replication at the endoplasmic reticulum is a shared replication strategy, despite different clinical outcomes. Although the protein products of these viruses have been studied in depth, how the RNA genomes operate inside human cells is poorly understood. Using comprehensive identification of RNA-binding proteins by mass spectrometry (ChIRP-MS), we took an RNA-centric viewpoint of flaviviral infection and identified several hundred proteins associated with both DENV and ZIKV genomic RNA in human cells. Genome-scale knockout screens assigned putative functional relevance to the RNA-protein interactions observed by ChIRP-MS. The endoplasmic-reticulum-localized RNA-binding proteins vigilin and ribosome-binding protein 1 directly bound viral RNA and each acted at distinct stages in the life cycle of flaviviruses. Thus, this versatile strategy can elucidate features of human biology that control the pathogenesis of clinically relevant viruses.
黄病毒，包括登革热病毒（DENV）和寨卡病毒（ZIKV），会导致严重的人类疾病。尽管临床结果不同，但在内质网上翻译病毒和病毒基因组复制所需的细胞因子是一种共同的复制策略。尽管这些病毒的蛋白质产物已经被深入研究，但 RNA 基因组在人类细胞内的运作方式仍知之甚少。我们通过质谱法（ChIRP-MS）对 RNA 结合蛋白进行全面鉴定，从 RNA 为中心的角度研究了黄病毒感染，并鉴定出数百种与人细胞中 DENV 和 ZIKV 基因组 RNA 相关的蛋白。基因组规模的敲除筛选将 ChIRP-MS 观察到的 RNA-蛋白相互作用赋予了推测的功能相关性。内质网定位的 RNA 结合蛋白 vigilantin 和核糖体结合蛋白 1 直接结合病毒 RNA，并且各自在黄病毒生命周期的不同阶段发挥作用。因此，这种多功能策略可以阐明控制具有临床相关性的病毒发病机制的人类生物学特征。
