Src Family Kinase Inhibitors Block Translation of Alphavirus Subgenomic mRNAs
Src Family Kinase Inhibitors Block Translation of Alphavirus Subgenomic mRNAs
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DOI：
10.1128/aac.02325-18
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发表时间：
2019-04-01
影响因子：
4.9
通讯作者：
Streblow, Daniel N.
中科院分区：
文献类型：
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作者：
Broeckel, Rebecca;Sarkar, Sanjay;Streblow, Daniel N.
甲病毒是节肢动物传播的RNA病毒，可引起人类关节痛、肌痛和脑炎。由于细胞激酶在甲病毒复制中的作用是未知的，我们分析了基孔肯雅病毒（CHIKV）感染成纤维细胞后宿主激酶丰度和磷酸化的动力学变化。基于本研究的结果，我们用靶向Src家族激酶（SFK）-磷脂酰肌醇3-激酶（PI 3 K）-AKT-mTORC信号传导通路的激酶抑制剂处理CHIKV感染的细胞。用SFK抑制剂处理细胞阻断了CHIKV以及多种其他甲病毒的复制，包括Mayaro病毒、O 'nyong-nyong病毒、Ross River病毒和委内瑞拉马脑炎病毒。剖析SFK抑制对甲病毒复制的影响，我们发现病毒结构蛋白水平显著降低，但病毒基因组和亚基因组RNA的合成不受影响。通过测量病毒RNA与多聚核糖体的结合，我们发现SFK抑制剂达沙替尼阻断了甲病毒亚基因组RNA的翻译。我们的研究结果表明SFK信号在甲病毒亚基因组RNA翻译和复制中的作用。靶向参与甲病毒复制的宿主因子代表了一种创新的、可能是范式转变的策略，用于探索CHIKV和其他甲病毒的复制，同时促进抗病毒治疗的开发。
Alphaviruses are arthropod-transmitted RNA viruses that can cause arthralgia, myalgia, and encephalitis in humans. Since the role of cellular kinases in alphavirus replication is unknown, we profiled kinetic changes in host kinase abundance and phosphorylation following chikungunya virus (CHIKV) infection of fibroblasts. Based upon the results of this study, we treated CHIKV-infected cells with kinase inhibitors targeting the Src family kinase (SFK)-phosphatidylinositol 3-kinase (PI3K)-AKT-mTORC signaling pathways. Treatment of cells with SFK inhibitors blocked the replication of CHIKV as well as multiple other alphaviruses, including Mayaro virus, O'nyong-nyong virus, Ross River virus, and Venezuelan equine encephalitis virus. Dissecting the effect of SFK inhibition on alphavirus replication, we found that viral structural protein levels were significantly reduced, but synthesis of viral genomic and subgenomic RNAs was unaffected. By measuring the association of viral RNA with polyribosomes, we found that the SFK inhibitor dasatinib blocks alphavirus subgenomic RNA translation. Our results demonstrate a role for SFK signaling in alphavirus subgenomic RNA translation and replication. Targeting host factors involved in alphavirus replication represents an innovative, perhaps paradigm-shifting, strategy for exploring the replication of CHIKV and other alphaviruses while promoting antiviral therapeutic development.