Rotavirus Genomic RNA Complex Forms via Specific RNA-RNA Interactions: Disruption of RNA Complex Inhibits Virus Infectivity.
Rotavirus Genomic RNA Complex Forms via Specific RNA-RNA Interactions: Disruption of RNA Complex Inhibits Virus Infectivity.
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DOI：
10.3390/v9070167
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发表时间：
2017-06-29
期刊：
影响因子：
--
通讯作者：
Roy P
中科院分区：
文献类型：
--
作者：
Fajardo T;Sung PY;Celma CC;Roy P
轮状病毒（RV）是呼肠孤病毒科（Reoviridae）的成员，引起儿童和婴儿感染，具有高发病率和死亡率。为了能够存活，病毒颗粒必须包装一组11个RNA片段。为了了解包装机制，在这里，我们在体外以不同的组合共合成了一组RNA片段，并通过两种替代方法进行检测：电泳迁移率变动试验（EMSA）和RNA珠下拉试验。我们发现，病毒正义RNA片段以特定的方式相互作用，形成RNA复合物，并且RNA-RNA相互作用遵循由小RV片段启动的顺序。此外，我们证明了RNA复合物被与短RNA序列互补的靶向特异性反义寡核苷酸（ORN）干扰，表明不同片段之间的RNA-RNA相互作用是序列特异性的。相同的抑制性ORN也具有抑制病毒复制的能力。结合体外和体内的数据推断，RNA-RNA相互作用和特定的复合物的形成是必不可少的排序不同的片段，可能之前，或期间，基因组包装。由于基因组组装是呼肠孤病毒科的普遍要求，这项工作提供了一种进一步了解RV和相关dsRNA（双链RNA）病毒的分选和包装机制的方法。
Rotavirus (RV), a member of the Reoviridae family, causes infection in children and infants, with high morbidity and mortality. To be viable, the virus particle must package a set of eleven RNA segments. In order to understand the packaging mechanism, here, we co-synthesized sets of RNA segments in vitro in different combinations and detected by two alternate methods: the electrophoretic mobility shift assay (EMSA) and the RNA-bead pull-down assay. We showed that viral positive-sense RNA segments interact with each other in a specific manner, forming RNA complexes, and that the RNA–RNA interactions followed a sequential order initiated by small RV segments. Further, we demonstrated that RNA complexes were perturbed by targeted specific antisense oligoribonucleotides (ORNs) complementary to short RNA sequences, indicating that the RNA–RNA interactions between different segments were sequence-specific. The same inhibitory ORNs also had the capability to inhibit virus replication. The combined in vitro and in vivo data inferred that RNA–RNA interactions and specific complex formation are essential for sorting different segments, possibly prior to, or during, genome packaging. As genome assembly is a universal requirement in the Reoviridae family, this work offers an approach towards a further understanding of the sorting and packaging mechanisms of RV and related dsRNA (double-stranded RNA) viruses.
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影响因子：
--
作者：
Fajardo T Jr;AlShaikhahmed K;Roy P
通讯作者：
Roy P
影响因子：
6.7
作者：
Fajardo T Jr;Sung PY;Roy P
通讯作者：
Roy P
影响因子：
7.7
作者：
Arias, CF;Isa, P;López, S
通讯作者：
López, S
影响因子：
5.4
作者：
Boyce, Mark;Celma, Cristina C. P.;Roy, Polly
通讯作者：
Roy, Polly
DOI：
10.1073/pnas.1220345110
发表时间：
2013-07-16
影响因子：
11.1
作者：
Periz, Javier;Celma, Cristina;Kapanidis, Achillefs N.
通讯作者：
Kapanidis, Achillefs N.