The conserved stem-loop II structure at the 3' untranslated region of Japanese encephalitis virus genome is required for the formation of subgenomic flaviviral RNA.

The conserved stem-loop II structure at the 3' untranslated region of Japanese encephalitis virus genome is required for the formation of subgenomic flaviviral RNA.
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DOI:
10.1371/journal.pone.0201250
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发表时间:
2018
期刊:
影响因子:
3.7
通讯作者:
Chang RY
Chang RY
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Chen YS;Fan YH;Tien CF;Yueh A;Chang RY

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黄病毒在感染细胞中积累丰富的亚基因组RNA(sfRNA)。据报道,sfRNA是由宿主5 '-至-3'核糖核酸外切酶XRN 1在3'非翻译区(UTR)的高度结构化RNA处停滞而产生的。尽管XRN 1对3 '端800-nt RNA的消化可以在体外停止在产生sfRNA的位置,但我们发现敲除XRN 1对日本脑炎病毒(JEV)感染细胞中sfRNA的积累没有影响。诱变研究揭示,3' UTR处的茎环II(SLII)是sfRNA积累所需的。根据体外RNA依赖性RNA聚合酶(RdRp)测定的结果,(-)10431-10566 RNA片段(其含有sfRNA转录的反基因组上的推定启动子)与RdRp蛋白结合并表现出强启动子活性。综上所述,我们的结果表明,JEV sfRNA可以最初转录,然后被XRN 1或其他未鉴定的核糖核酸外切酶修剪。
Flaviviruses accumulate abundant subgenomic RNA (sfRNA) in infected cells. It has been reported that sfRNA results from stalling of host 5’-to-3’ exoribonuclease XRN1 at the highly structured RNA of the 3’ untranslated region (UTR). Although XRN1 digestion of a 3’-terminal 800-nt RNA could stall at a position to generate the sfRNA in vitro, we found that knocking out XRN1 had no effect on the accumulation of sfRNA in Japanese encephalitis virus (JEV) infected cells. Mutagenesis studies revealed that the stemloop II (SLII) at the 3’ UTR is required for the accumulation of sfRNA. According to the results of an in vitro RNA-dependent RNA polymerase (RdRp) assay, the (-)10431-10566 RNA fragment, containing the putative promoter on the antigenome for the sfRNA transcription, binds to RdRp protein and exhibits a strong promoter activity. Taken together, our results indicate that the JEV sfRNA could be transcribed initially and then be trimmed by XRN1 or other unidentified exoribonucleases.
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