Functional Mapping of AGO-Associated Zika Virus-Derived Small Interfering RNAs in Neural Stem Cells.
Functional Mapping of AGO-Associated Zika Virus-Derived Small Interfering RNAs in Neural Stem Cells.
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AGO相关寨卡病毒衍生的小干扰RNA在神经干细胞中的功能定位。
DOI：
10.3389/fcimb.2021.628887
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发表时间：
2021
影响因子：
5.7
通讯作者：
Zhao Z
中科院分区：
文献类型：
--
作者：
Zeng J;Luo Z;Dong S;Xie X;Liang X;Yan Y;Liang Q;Zhao Z
通过对包括寨卡病毒（ZIKV）在内的病毒感染细胞进行直接深度RNA测序，已从几种病毒基因组中鉴定出病毒干扰RNA （viRNA）。一旦由核糖核酸内切酶Dicer产生，viRNAs就被装载到RNA诱导沉默复合物（RISCs）的Argonaute （AGO）家族蛋白上，与它们的RNA靶标配对并启动靶基因的切割。然而，功能性ZIKV病毒及其病毒RNA靶标的身份在很大程度上仍然未知。我们最近的研究表明，ZIKV衣壳蛋白与Dicer相互作用并拮抗其核糖核酸内切酶活性，这需要它在第41个氨基酸上的组氨酸残基。因此，经过改造的ZIKV-H41R功能缺失（LOF）突变病毒不再抑制Dicer酶活性，也不再抑制NSCs中的miRNA生物发生。通过结合ago相关RNA测序、ZIKV感染的人神经干细胞（NSCs）的深度测序分析和miRanda靶标扫描，我们在NSCs中定义了29个ZIKV衍生的viRNA谱，并建立了viRNA与病毒靶标之间复杂的相互作用网络。更重要的是，我们发现由ZIKV mRNA产生的viRNA依赖于Dicer功能，并且是NSCs中ZIKV毒力的限制因素。结果，由ZIKV-H41R病毒感染的NSCs产生的virna水平要高得多。因此，我们将viRNAs映射到它们的RNA靶标上，为进一步研究viRNAs如何在抗病毒机制中发挥作用，以及其他未知的生物学功能铺平了道路。
Viral interfering RNA (viRNA) has been identified from several viral genomes via directly deep RNA sequencing of the virus-infected cells, including zika virus (ZIKV). Once produced by endoribonuclease Dicer, viRNAs are loaded onto the Argonaute (AGO) family proteins of the RNA-induced silencing complexes (RISCs) to pair with their RNA targets and initiate the cleavage of target genes. However, the identities of functional ZIKV viRNAs and their viral RNA targets remain largely unknown. Our recent study has shown that ZIKV capsid protein interacted with Dicer and antagonized its endoribonuclease activity, which requires its histidine residue at the 41st amino acid. Accordingly, the engineered ZIKV-H41R loss-of-function (LOF) mutant virus no longer suppresses Dicer enzymatic activity nor inhibits miRNA biogenesis in NSCs. By combining AGO-associated RNA sequencing, deep sequencing analysis in ZIKV-infected human neural stem cells (NSCs), and miRanda target scanning, we defined 29 ZIKV derived viRNA profiles in NSCs, and established a complex interaction network between the viRNAs and their viral targets. More importantly, we found that viRNA production from the ZIKV mRNA is dependent on Dicer function and is a limiting factor for ZIKV virulence in NSCs. As a result, much higher levels of viRNAs generated from the ZIKV-H41R virus-infected NSCs. Therefore, our mapping of viRNAs to their RNA targets paves a way to further investigate how viRNAs play the role in anti-viral mechanisms, and perhaps other unknown biological functions.