Interferon-Stimulated Gene (ISG)-Expression Screening Reveals the Specific Antibunyaviral Activity of ISG20.
Interferon-Stimulated Gene (ISG)-Expression Screening Reveals the Specific Antibunyaviral Activity of ISG20.
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DOI：
10.1128/jvi.02140-17
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发表时间：
2018-07-01
影响因子：
5.4
通讯作者：
Wilson SJ
中科院分区：
文献类型：
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作者：
Feng J;Wickenhagen A;Turnbull ML;Rezelj VV;Kreher F;Tilston-Lunel NL;Slack GS;Brennan B;Koudriakova E;Shaw AE;Rihn SJ;Rice CM;Bieniasz PD;Elliott RM;Shi X;Wilson SJ
布尼亚病毒对人类健康、繁荣和粮食安全构成重大威胁。在病毒感染的反应中，干扰素（ifn）上调数百种干扰素刺激基因（isg）的表达，其累积作用可以有效抑制兔病毒的复制。我们使用基于流式细胞术的方法筛选了来自人类和恒河猴的约500个独特的isg抑制Bunyamwera正布尼亚病毒（BUNV）复制的能力，Bunyamwera正布尼亚病毒（BUNV）是布尼亚病毒科和布尼亚病毒目的原型。候选候选具有抗布尼亚病毒活性，进一步检查使用一组不同的布尼亚病毒。有趣的是，一种候选药物ISG20对大多数来自环布尼亚病毒科、汉坦病毒科和奈罗病毒科的病毒表现出有效的抗布尼亚病毒活性，而白蛉病毒（Phenuiviridae）基本上没有受到抑制。与已知ISG20靶向的其他病毒类似，ISG20的抗病毒活性依赖于其功能性rna酶活性。通过传染性病毒样颗粒（VLP）测定（基于BUNV微小基因组系统），我们证实了ISG20强烈抑制了所有3个病毒片段的基因表达。通过体外进化，我们生成了一个具有ISG20抗性的BUNV，并绘制了ISG20敏感性/抗性的决定因素。综合所有数据，我们报道ISG20是一种广泛而有效的抗布尼亚病毒因子，但一些布尼亚病毒具有显著的ISG20抗性。因此，ISG20敏感性/耐药性可能影响布尼亚病毒的发病机制，其中许多是具有临床或兽医意义的新兴病毒。布尼亚病毒有数百种，其中许多在人类和牲畜中引起危及生命的急性疾病。干扰素（IFN）系统是先天免疫的关键组成部分，I型干扰素在体内和体外都限制了布尼亚病毒的繁殖。I型IFN信号导致数百种IFN刺激基因（isg）的上调，其协同作用产生“抗病毒状态”。尽管干扰素在限制布尼亚病毒的复制和发病机制方面至关重要，但对于哪些干扰素抑制布尼亚病毒，我们仍然知之甚少。通过isg表达筛选，研究人员检测了约500种独特的isg抑制布尼亚病毒（Bunyamwera orthobunyavirus， BUNV）的能力。利用这种方法，我们发现干扰素刺激的外切酶ISG20是一种有效的BUNV抑制剂。有趣的是，ISG20具有高度选择性的抗布尼亚病毒活性，多种布尼亚病毒被有效抑制，而一些病毒基本上逃脱了抑制。我们推测，一些布尼亚病毒逃避ISG20的能力可能影响其发病机制。
Bunyaviruses pose a significant threat to human health, prosperity, and food security. In response to viral infections, interferons (IFNs) upregulate the expression of hundreds of interferon-stimulated genes (ISGs), whose cumulative action can potently inhibit the replication of bunyaviruses. We used a flow cytometry-based method to screen the ability of ∼500 unique ISGs from humans and rhesus macaques to inhibit the replication of Bunyamwera orthobunyavirus (BUNV), the prototype of both the Peribunyaviridae family and the Bunyavirales order. Candidates possessing antibunyaviral activity were further examined using a panel of divergent bunyaviruses. Interestingly, one candidate, ISG20, exhibited potent antibunyaviral activity against most viruses examined from the Peribunyaviridae, Hantaviridae, and Nairoviridae families, whereas phleboviruses (Phenuiviridae) largely escaped inhibition. Similar to the case against other viruses known to be targeted by ISG20, the antibunyaviral activity of ISG20 is dependent upon its functional RNase activity. Through use of an infectious virus-like particle (VLP) assay (based on the BUNV minigenome system), we confirmed that gene expression from all 3 viral segments is strongly inhibited by ISG20. Using in vitro evolution, we generated a substantially ISG20-resistant BUNV and mapped the determinants of ISG20 sensitivity/resistance. Taking all the data together, we report that ISG20 is a broad and potent antibunyaviral factor but that some bunyaviruses are remarkably ISG20 resistant. Thus, ISG20 sensitivity/resistance may influence the pathogenesis of bunyaviruses, many of which are emerging viruses of clinical or veterinary significance. IMPORTANCE There are hundreds of bunyaviruses, many of which cause life-threatening acute diseases in humans and livestock. The interferon (IFN) system is a key component of innate immunity, and type I IFNs limit bunyaviral propagation both in vitro and in vivo. Type I IFN signaling results in the upregulation of hundreds of IFN-stimulated genes (ISGs), whose concerted action generates an “antiviral state.” Although IFNs are critical in limiting bunyaviral replication and pathogenesis, much is still unknown about which ISGs inhibit bunyaviruses. Using ISG-expression screening, we examined the ability of ∼500 unique ISGs to inhibit Bunyamwera orthobunyavirus (BUNV), the prototypical bunyavirus. Using this approach, we identified ISG20, an interferon-stimulated exonuclease, as a potent inhibitor of BUNV. Interestingly, ISG20 possesses highly selective antibunyaviral activity, with multiple bunyaviruses being potently inhibited while some largely escape inhibition. We speculate that the ability of some bunyaviruses to escape ISG20 may influence their pathogenesis.