A general strategy to inhibiting viral-1 frameshifting based on upstream attenuation duplex formation
A general strategy to inhibiting viral-1 frameshifting based on upstream attenuation duplex formation
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DOI：
10.1093/nar/gkv1307
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发表时间：
2016-01-08
影响因子：
14.9
通讯作者：
Chang, Kung-Yao
中科院分区：
文献类型：
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作者：
Hu, Hao-Teng;Cho, Che-Pei;Chang, Kung-Yao
由于包括人类免疫缺陷病毒(HIV)和冠状病毒在内的许多人类病毒病原体依赖于-1核糖体移码(PRF)进行最佳繁殖，因此病毒-1程序性核糖体移码(PRF)作为一种潜在的抗病毒靶点已引起人们的兴趣。高效的真核细胞-1PRF需要一个位于移码位点下游的最佳刺激子结构，针对病毒-1PRF刺激子的不同策略已经被开发出来。然而，获取特定的PRF-1刺激物信息是抗击中东呼吸综合征冠状病毒(MERS-CoV)等新出现的流行病毒病原体的瓶颈。最近，一个位于移码位点上游的RNA发夹被证明是一个顺式元件，可以减弱-1PRF，但机制未知。在这里，我们表明，通过在移码位点上游对其互补的mRNA序列进行反义退火，形成的上游双链可以取代上游发夹，有效地衰减-1PRF。这一发现表明，近端上游双链的形成是导致-1\f25 PRF-1衰减的主要决定因素，并提供了机械性见解。此外，反义介导的上游双链方法下调了不同的-1PRF刺激子刺激的-1PRF，包括MERS-CoV的刺激因子，这表明一旦有新出现的人类冠状病毒的序列信息，它就有可能作为一种强大的手段来评估病毒-1PRF的抑制作用。
Viral -1 programmed ribosomal frameshifting (PRF) as a potential antiviral target has attracted interest because many human viral pathogens, including human immunodeficiency virus (HIV) and coronaviruses, rely on -1 PRF for optimal propagation. Efficient eukaryotic -1 PRF requires an optimally placed stimulator structure downstream of the frameshifting site and different strategies targeting viral -1 PRF stimulators have been developed. However, accessing particular -1 PRF stimulator information represents a bottle-neck in combating the emerging epidemic viral pathogens such as Middle East respiratory syndrome coronavirus (MERS-CoV). Recently, an RNA hairpin upstream of frameshifting site was shown to act as a cis-element to attenuate -1 PRF with mechanism unknown. Here, we show that an upstream duplex formed in-trans, by annealing an antisense to its complementary mRNA sequence upstream of frameshifting site, can replace an upstream hairpin to attenuate -1 PRF efficiently. This finding indicates that the formation of a proximal upstream duplex is the main determining factor responsible for -1 PRF attenuation and provides mechanistic insight. Additionally, the antisense-mediated upstream duplex approach downregulates -1 PRF stimulated by distinct -1 PRF stimulators, including those of MERS-CoV, suggesting its general application potential as a robust means to evaluating viral -1 PRF inhibition as soon as the sequence information of an emerging human coronavirus is available.