Conserved RNA secondary structure in Cherry virus A 5'-UTR associated with translation regulation.
Conserved RNA secondary structure in Cherry virus A 5'-UTR associated with translation regulation.
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DOI：
10.1186/s12985-022-01824-z
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发表时间：
2022-05-26
期刊：
影响因子：
4.8
通讯作者：
Yuan, Xuefeng
中科院分区：
文献类型：
--
作者：
Wang, Deya;Yang, Chen;Deng, Yanmei;Cao, Xue;Xu, Wei;Han, Zishuo;Li, Qingliang;Yang, Yang;Yuan, Xuefeng
病毒基因组5′非翻译区（UTR）或3′非翻译区中具有多种结构的顺式作用RNA元件在病毒翻译过程中发挥着关键作用。樱桃病毒A（CVA）是β-柔线病毒科毛形病毒属的成员。其基因组为正义单链RNA，约7400个核苷酸（nt）。CVA的5′非翻译区长度约为100 nt；然而，这一较长非翻译区的功能尚未见报道。
对75条CVA序列进行了分子及系统发育分析，这些序列可分为四组，并对四种CVA 5′非翻译区类型的RNA二级结构进行了预测。随后，将这四种CVA 5′非翻译区类型分别插入萤火虫荧光素酶报告基因FLuc（FLuc）的上游，利用小麦胚芽提取物（WGE）对相应转录本进行体外翻译评估。接着，进行内联结构探测以揭示CVA 5′非翻译区中保守的RNA结构。
这四种CVA 5′非翻译区类型似乎具有保守的RNA结构，含有这四种CVA 5′非翻译区类型的FLuc构建体使FLuc的翻译水平提高了2 - 3倍，表明其具有较弱的翻译增强活性。CVA 5′非翻译区的突变抑制了翻译，这表明保守的RNA结构对其功能很重要。
通过对不同CVA分离株的结构进化分析鉴定出了保守的RNA二级结构，发现其对翻译具有调控作用。
A variety of cis-acting RNA elements with structures in the 5′- or 3′-untranslated region (UTR) of viral genomes play key roles in viral translation. Cherry virus A (CVA) is a member of the genus Capillovirus in the family Betaflexiviridae. It has a positive single-stranded RNA genome of ~ 7400 nucleotides (nt). The length of the CVA 5′-UTR is ~ 100 nt; however, the function of this long UTR has not yet been reported. Molecular and phylogenetic analyses were performed on 75 CVA sequences, which could be divided into four groups, and the RNA secondary structure was predicted in four CVA 5′-UTR types. These four CVA 5′-UTR types were then inserted upstream of the firefly luciferase reporter gene FLuc (FLuc), and in vitro translation of the corresponding transcripts was evaluated using wheat germ extract (WGE). Then, in-line structure probing was performed to reveal the conserved RNA structures in CVA-5′UTR. The four CVA 5′-UTR types appeared to have a conserved RNA structure, and the FLuc construct containing these four CVA 5′-UTR types increased the translation of FLuc by 2–3 folds, suggesting weak translation enhancement activity. Mutations in CVA 5′-UTR suppressed translation, suggesting that the conserved RNA structure was important for function. The conserved RNA secondary structure was identified by structural evolution analysis of different CVA isolates and was found to regulate translation.
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