RNA elements required for RNA recombination function as replication enhancers in vitro and in vivo in a plus-strand RNA virus
RNA elements required for RNA recombination function as replication enhancers in vitro and in vivo in a plus-strand RNA virus
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DOI：
10.1093/emboj/18.20.5653
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发表时间：
1999-10-15
期刊：
影响因子：
11.4
通讯作者：
Simon, AE
中科院分区：
文献类型：
--
作者：
Nagy, PD;Pogany, J;Simon, AE
RNA 复制需要顺式作用元件来募集病毒 RNA 依赖性 RNA 聚合酶 (RdRp) 并促进互补链合成的从头启动。发夹是萝卜皱病毒 (TCV) 和卫星 (sat)-RNAC（一种与 TCV 感染相关的寄生 RNA）基因组 RNA 中重组的热点，在使用部分纯化的 TCV RdRp 进行的体外测定中，可将下游启动子序列的 RNA 合成刺激 10 倍。人工发夹对转录具有抑制作用。当将天然茎环结构插入珍珠状积累的 sat-RNA 中时，单细胞中的 RNA 积累增加了 5 至 10 倍。茎环结构对RNA复制的影响是相加的，插入三个茎环RNA元件可以最大程度地增加sat-RNA的积累（25倍）。这些茎环结构不会影响体内 RNA 的稳定性，但可能有助于将 RdRp 招募到模板上。
RNA replication requires cis-acting elements to recruit the viral RNA-dependent RNA polymerase (RdRp) and facilitate de novo initiation of complementary strand synthesis. Hairpins that are hot spots for recombination in the genomic RNA of turnip crinkle virus (TCV) and satellite (sat)-RNAC, a parasitic RNA associated with TCV infections, stimulate RNA synthesis 10-fold from a downstream promoter sequence in an in vitro assay using partially purified TCV RdRp, Artificial hairpins had an inhibitory effect on transcription. RNA accumulation in single cells was enhanced 5- to 10-fold when the natural stem-loop structures were inserted into a pearly accumulating sat-RNA. The effect of the stem-loop structures on RNA replication was additive, with insertion of three stem-loop RNA elements increasing sat-RNA accumulation to the greatest extent (25-fold). These stem-loop structures do not influence the stability of the RNAs in vivo, but may serve to recruit the RdRp to the template.