Department of Microbiology, Immunology, and Molecular Genetics, University of Texas Health at San Antonio, San Antonio, TX 78229.
Department of Biology, Viral Immunology Center, Georgia State University, Atlanta, GA 30303;
Proc Natl Acad Sci U S A. 2020 Aug 18;117(33):20109-20116. doi: 10.1073/pnas.1921315117. Epub 2020 Aug 3.
Herpesviruses are ubiquitous human pathogens that cause a wide range of health complications. Currently, there is an incomplete understanding of cellular factors that contribute to herpesvirus infection. Here, we report an antiviral necroptosis-based genetic screen to identify novel host cell factors required for infection with the β-herpesvirus murine cytomegalovirus (MCMV). Our genome-wide CRISPR-based screen harnessed the capacity of herpesvirus mutants that trigger antiviral necroptotic cell death upon early viral gene expression. Vascular endothelial growth factor (VEGF) and semaphorin-binding receptor Neuropilin-1 (Nrp-1) emerge as crucial determinants of MCMV infection. We find that elimination of Nrp-1 impairs early viral gene expression and reduces infection rates in endothelial cells, fibroblasts, and macrophages. Furthermore, preincubation of virus with soluble Nrp-1 dramatically inhibits infection by reducing virus attachment. Thus, Nrp-1 is a key determinant of the initial phase of MCMV infection.
疱疹病毒是普遍存在的人类病原体,可引起多种健康并发症。目前,人们对有助于疱疹病毒感染的细胞因子了解不完整。在这里,我们报告了一种基于抗病毒坏死性细胞凋亡的遗传筛选,以鉴定感染β疱疹病毒鼠巨细胞病毒(MCMV)所需的新型宿主细胞因子。我们基于全基因组 CRISPR 的筛选利用了疱疹病毒突变体的能力,这些突变体在早期病毒基因表达时引发抗病毒坏死性细胞死亡。血管内皮生长因子(VEGF)和神经纤毛蛋白结合受体 Neuropilin-1(Nrp-1)成为 MCMV 感染的关键决定因素。我们发现,消除 Nrp-1 会损害早期病毒基因表达,并降低内皮细胞、成纤维细胞和巨噬细胞中的感染率。此外,用可溶性 Nrp-1 预先孵育病毒可通过减少病毒附着来显著抑制感染。因此,Nrp-1 是 MCMV 感染初始阶段的关键决定因素。