手,脚,和口蹄疫(HFMD)是婴儿和儿童的常见传染病,尤其是五岁以下的人。EV-A71是引起HFMD的常见病原体,也是导致严重或致命HFMD的主要病原体,以神经系统并发症为特征。然而,EV-A71发病机制的基本机制仍不清楚.在这份报告中,我们使用蛋白质组学和磷酸化蛋白质组学方法来表征EV-A71感染的人神经母细胞瘤SK-N-SH细胞的蛋白质组和磷酸蛋白质组特征。成功定量了超过7744个宿主蛋白和10069个磷酸化修饰位点。其中,在EV-A71感染过程中,974个蛋白质和3648个磷酸化修饰位点受到显着调节。KEGG(京都基因和基因组百科全书)途径分析显示,EV-A71改变了细胞生物学过程,包括蛋白质合成,SK-N-SH细胞中的RNA剪接和代谢。值得注意的是,基于EV-A71感染期间激酶上调的预测,我们确定了FDA批准的特异性激酶抑制剂,与ceralasertib,博舒替尼,黄素单核苷酸,米诺环素,pimasertib和乙酰半胱氨酸抑制EV-A71感染。最后,EV-A71蛋白在感染过程中被磷酸化,观察到一个位点(3D聚合酶上的S184)对病毒复制至关重要,因为S184A突变敲除了病毒复制。结果提高了我们对宿主对神经母细胞瘤细胞EV-A71感染反应的理解,并为开发抗EV-A71策略提供了潜在的靶标。
Hand, foot, and mouth disease (HFMD) is a common infectious disease in infants and children, especially those under five years of age. EV-A71 is a common pathogen that causes HFMD and the primary pathogen leading to severe or fatal HFMD, which is characterized by neurological complications. However, the underlying mechanisms of EV-A71 pathogenesis remain largely unknown. In this report, we used proteomic and phosphorylated proteomic methods to characterize the proteome and phosphoproteome profiles of EV-A71-infected human neuroblastoma SK-N-SH cells. More than 7744 host proteins and 10069 phosphorylation modification sites were successfully quantified. Among them, 974 proteins and 3648 phosphorylation modification sites were regulated significantly during EV-A71 infection. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis revealed that EV-A71 altered cell biological processes, including protein synthesis, RNA splicing and metabolism in SK-N-SH cells. Notably, based on the prediction of upregulated kinases during EV-A71 infection, we identified specific kinase inhibitors approved by the FDA, with ceralasertib, bosutinib, flavin mononucleotide, minocycline, pimasertib and acetylcysteine inhibiting EV-A71 infection. Finally, EV-A71 proteins were found to be phosphorylated during infection, with one site (S184 on 3D polymerase) observed to be crucial for viral replication because a S184A mutation knocked out viral replication. The results improve our understanding of the host response to EV-A71 infection of neuroblastoma cells and provide potential targets for developing anti-EV-A71 strategies.