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miR-122 does not modulate the elongation phase of hepatitis C virus RNA synthesis in isolated replicase complexes.

机译:miR-122不会调节分离的复制酶复合物中丙型肝炎病毒RNA合成的延长阶段。

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摘要

miR-122 is an abundant, liver-specific microRNA that is required for efficient amplification of hepatitis C virus (HCV) RNA. Recent studies with a miR-122-specific locked nucleic acid antagomir have shown it to be an important host target for therapeutic intervention. However, considerable controversy exists concerning the mechanisms underlying the dependence of HCV replication on miR-122. We studied the impact of miR-122 on the rate of [(32)P]-incorporation into positive-strand viral RNA by membrane-bound replicase complexes isolated from cells containing HCV RNA replicons. [(32)P]-incorporation in this cell-free system represents primarily the elongation phase of RNA synthesis, with little or no de novo initiation, and was not affected by the addition of either excess miR-122 or a miR-122-specific antisense oligonucleotide that suppresses replication in vivo. We also found no evidence that detectable quantities of miR-122 are specifically associated with replicase complexes in vivo. These results are consistent with miR-122 acting at an alternative step in the viral life cycle, promoting cap-independent viral translation, enhancing viral RNA stability, or facilitating de novo initiation of viral RNA synthesis.
机译:miR-122是丰富的肝特异性microRNA,是有效扩增丙型肝炎病毒(HCV)RNA所必需的。最近对miR-122特异性锁定核酸antagomir的研究表明,它是治疗干预的重要宿主靶标。然而,关于HCV复制对miR-122的依赖性所依据的机制存在相当大的争议。我们研究了膜隔离复制酶复合物从含有HCV RNA复制子的细胞中分离出来对miR-122对[(32)P]掺入正链病毒RNA的速率的影响。 [(32)P]掺入此无细胞系统主要代表RNA合成的延伸阶段,几乎没有或没有从头开始,并且不受添加过量miR-122或miR-122-的影响抑制体内复制的特定反义寡核苷酸。我们还没有发现证据表明可检测量的miR-122与体内复制酶复合物特别相关。这些结果与miR-122在病毒生命周期的另一个步骤起作用,促进不依赖帽的病毒翻译,增强病毒RNA稳定性或促进从头开始病毒RNA合成相一致。

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