Positive strand RNA viruses differ in the constraints they place on the folding of their negative strand.
Positive strand RNA viruses differ in the constraints they place on the folding of their negative strand.
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DOI：
10.1261/rna.079125.122
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发表时间：
2022-10
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--
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正链RNA病毒的基因组复制需要产生互补的负链RNA，其用作合成更多正链子代的模板。结构RNA元件对于基因组复制是重要的，但是虽然它们在正链中很容易观察到，但它们在负链中存在的证据更加有限。我们假设这是由于病毒允许后者RNA采用结构折叠的能力不同。为了研究这一点，将核酶引入到不同病毒构建体的负链中;预期如果RNA折叠发生，将观察到负链切割和复制抑制。事实上，这就是丙型肝炎病毒（HCV）和猫杯状病毒（FCV）构建体所发生的情况。然而，对基孔肯雅病毒（CHIKV）、人鼻病毒（HRV）、戊型肝炎病毒（HEV）和黄热病病毒（YFV）构建体几乎没有观察到影响。证明负链中的切割减少是由于与正链形成双链体。有趣的是，含核酶的RNA在体外由HCV聚合酶产生时也保持完整，这也是由于双链体的形成。总体而言，我们的研究结果表明，在不同的正链RNA病毒之间的负链折叠的构象约束有重要的差异。
Genome replication of positive strand RNA viruses requires the production of a complementary negative strand RNA that serves as a template for synthesis of more positive strand progeny. Structural RNA elements are important for genome replication, but while they are readily observed in the positive strand, evidence of their existence in the negative strand is more limited. We hypothesized that this was due to viruses differing in their capacity to allow this latter RNA to adopt structural folds. To investigate this, ribozymes were introduced into the negative strand of different viral constructs; the expectation being that if RNA folding occurred, negative strand cleavage and suppression of replication would be seen. Indeed, this was what happened with hepatitis C virus (HCV) and feline calicivirus (FCV) constructs. However, little or no impact was observed for chikungunya virus (CHIKV), human rhinovirus (HRV), hepatitis E virus (HEV), and yellow fever virus (YFV) constructs. Reduced cleavage in the negative strand proved to be due to duplex formation with the positive strand. Interestingly, ribozyme-containing RNAs also remained intact when produced in vitro by the HCV polymerase, again due to duplex formation. Overall, our results show that there are important differences in the conformational constraints imposed on the folding of the negative strand between different positive strand RNA viruses.
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