Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Südufer 10, 17493 Greifswald-Insel Riems, Germany.
Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Südufer 10, 17493 Greifswald-Insel Riems, Germany.
Viruses. 2023 May 30;15(6):1283. doi: 10.3390/v15061283.
African swine fever is a viral disease of swine caused by the African swine fever virus (ASFV). Currently, ASFV is spreading over the Eurasian continent and threatening global pig husbandry. One viral strategy to undermine an efficient host cell response is to establish a global shutoff of host protein synthesis. This shutoff has been observed in ASFV-infected cultured cells using two-dimensional electrophoresis combined with metabolic radioactive labeling. However, it remained unclear if this shutoff was selective for certain host proteins. Here, we characterized ASFV-induced shutoff in porcine macrophages by measurement of relative protein synthesis rates using a mass spectrometric approach based on stable isotope labeling with amino acids in cell culture (SILAC). The impact of ASFV infection on the synthesis of >2000 individual host proteins showed a high degree of variability, ranging from complete shutoff to a strong induction of proteins that are absent from naïve cells. GO-term enrichment analysis revealed that the most effective shutoff was observed for proteins related to RNA metabolism, while typical representatives of the innate immune system were strongly induced after infection. This experimental setup is suitable to quantify a virion-induced host shutoff (vhs) after infection with different viruses.
非洲猪瘟是一种由非洲猪瘟病毒（ASFV）引起的猪病毒性疾病。目前，ASF 在欧亚大陆蔓延，威胁着全球养猪业。病毒破坏有效宿主细胞反应的一种策略是建立宿主蛋白合成的全局关闭。在用二维电泳结合代谢放射性标记的方法研究 ASFV 感染的培养细胞时，已经观察到这种关闭。然而，目前尚不清楚这种关闭是否对某些宿主蛋白具有选择性。在这里，我们通过使用基于稳定同位素标记的细胞培养中的氨基酸（SILAC）的质谱方法测量相对蛋白合成率，来表征猪巨噬细胞中 ASFV 诱导的关闭。ASFV 感染对超过 2000 种单个宿主蛋白的合成的影响表现出高度的可变性，范围从完全关闭到对幼稚细胞中不存在的蛋白的强烈诱导。GO 术语富集分析表明，观察到的最有效的关闭是与 RNA 代谢相关的蛋白，而感染后先天免疫系统的典型代表被强烈诱导。该实验方案适用于在感染不同病毒后量化病毒诱导的宿主关闭（vhs）。
