Mechanistic insight into the efficient packaging of antigenomic S RNA into Rift Valley fever virus particles.
Mechanistic insight into the efficient packaging of antigenomic S RNA into Rift Valley fever virus particles.
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机械洞察力洞察抗原学S RNA在裂谷发烧病毒颗粒中的有效包装。
DOI：
10.3389/fcimb.2023.1132757
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发表时间：
2023
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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作者：
裂谷热病毒（RVFV）是一种布尼亚病毒，具有单链负义三片段RNA基因组，由L、M和S RNA组成。传染性病毒粒子携带两种包膜糖蛋白，Gn和Gc，以及由封装病毒RNA片段组成的核糖核蛋白复合物。抗基因组S RNA作为编码非结构蛋白NSs（一种干扰素拮抗剂）的mRNA的模板，也被有效地包装到RVFV颗粒中。Gn与病毒核糖核蛋白复合物之间的相互作用，包括Gn与病毒RNA的直接结合，驱动病毒RNA包装进入RVFV颗粒。为了了解RVFV中有效的抗基因组S RNA包装机制，我们通过使用抗Gn抗体对RVFV感染的细胞裂解物进行紫外交联和免疫沉淀，然后进行高通量测序分析（CLIP-seq分析），确定了病毒RNA中与Gn直接相互作用的区域。我们的数据表明，在RVFV RNA中存在多个gn结合位点，包括在抗基因组S RNA的3 '非编码区内的一个突出的gn结合位点。我们发现，在RVFV突变体中，抗基因组S RNA的有效包装在3 '非编码区缺乏部分突出的gn结合位点时被取消。此外，突变RVFV在感染后触发干扰素-β mRNA表达的早期诱导，而不是亲本RVFV。这些数据表明，Gn与抗基因组S RNA的3 '非编码区RNA元件的直接结合促进了抗基因组S RNA有效地包装成病毒粒子。此外，在RNA元件的驱动下，抗基因组S RNA有效地包装到RVFV颗粒中，促进了感染后立即合成编码NSs的病毒mRNA，从而抑制了干扰素-β mRNA的表达。
Rift Valley fever virus (RVFV), a bunyavirus, has a single-stranded, negative-sense tri-segmented RNA genome, consisting of L, M and S RNAs. An infectious virion carries two envelope glycoproteins, Gn and Gc, along with ribonucleoprotein complexes composed of encapsidated viral RNA segments. The antigenomic S RNA, which serves as the template of the mRNA encoding a nonstructural protein, NSs, an interferon antagonist, is also efficiently packaged into RVFV particles. An interaction between Gn and viral ribonucleoprotein complexes, including the direct binding of Gn to viral RNAs, drives viral RNA packaging into RVFV particles. To understand the mechanism of efficient antigenomic S RNA packaging in RVFV, we identified the regions in viral RNAs that directly interact with Gn by performing UV-crosslinking and immunoprecipitation of RVFV-infected cell lysates with anti-Gn antibody followed by high-throughput sequencing analysis (CLIP-seq analysis). Our data suggested the presence of multiple Gn-binding sites in RVFV RNAs, including a prominent Gn-binding site within the 3’ noncoding region of the antigenomic S RNA. We found that the efficient packaging of antigenomic S RNA was abrogated in a RVFV mutant lacking a part of this prominent Gn-binding site within the 3’ noncoding region. Also, the mutant RVFV, but not the parental RVFV, triggered the early induction of interferon-β mRNA expression after infection. These data suggest that the direct binding of Gn to the RNA element within the 3’ noncoding region of the antigenomic S RNA promoted the efficient packaging of antigenomic S RNA into virions. Furthermore, the efficient packaging of antigenomic S RNA into RVFV particles, driven by the RNA element, facilitated the synthesis of viral mRNA encoding NSs immediately after infection, resulting in the suppression of interferon-β mRNA expression.
DOI：
10.3390/v13122417
发表时间：
2021-12-02
期刊：
Viruses
影响因子：
--
作者：
Hayashi M;Schultz EP;Lanchy JM;Lodmell JS
通讯作者：
Lodmell JS