Subgenomic negative-strand RNA function during mouse hepatitis virus infection
Subgenomic negative-strand RNA function during mouse hepatitis virus infection
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DOI：
10.1128/jvi.74.9.4039-4046.2000
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发表时间：
2000-05-01
影响因子：
5.4
通讯作者：
Yount, B
中科院分区：
文献类型：
--
作者：
Baric, RS;Yount, B
小鼠肝炎病毒（MHV）感染的细胞含有全长和亚基因组长度的正链和负链RNA。亚基因组负链RNA的起源和功能一直存在争议。在这份报告中，我们证明了亚基因组负链的合成和摩尔比在替代宿主细胞中是相似的，这表明这些RNA作为正链合成的重要介质发挥作用。使用动力学标记实验，我们表明，全长和亚基因组长度的复制形式的RNA迅速积累，然后饱和的标签，这表明亚基因组长度的负链是正链合成的主要介质。使用放线菌酮，它优先抑制负链，并在较小程度上的正链合成，我们表明，放线菌酮治疗同样抑制全长和亚基因组长度的负链合成。重要的是，治疗后，先前转录的负链保持在转录活性复合物中，即使在没有新的负链合成的情况下。这些结果表明，亚基因组长度的负链是MHV感染过程中正链合成的主要模板。
Mouse hepatitis virus (MHV)-infected cells contain full-length and subgenomic-length positive- and negative-strand RNAs. The origin and function of the subgenomic negative-strand RNAs is controversial. In this report we demonstrate that the synthesis and molar ratios of subgenomic negative strands are similar in alternative host cells, suggesting that these RNAs function as important mediators of positive-strand synthesis. Using kinetic labeling experiments, we show that the full-length and subgenomic-length replicative form RNAs rapidly accumulate and then saturate with label, suggesting that the subgenomic-length negative strands are the principal mediators of positive-strand synthesis. Using cycloheximide, which preferentially inhibits negative-strand and to a lesser extent positive-strand synthesis, we demonstrate that cycloheximide treatment equally inhibits full-length and subgenomic-length negative-strand synthesis. Importantly, following treatment, previously transcribed negative strands remain in transcriptionally active complexes even in the absence of new negative-strand synthesis. These findings indicate that the subgenomic-length negative strands are the principal templates of positive-strand synthesis during MHV infection.