miR-122 与 5' 外切核酸酶 Xrn1 在调控丙型肝炎病毒复制中的竞争和非竞争活性。

Competing and noncompeting activities of miR-122 and the 5' exonuclease Xrn1 in regulation of hepatitis C virus replication.

机构信息

Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7292, USA.

出版信息

Proc Natl Acad Sci U S A. 2013 Jan 29;110(5):1881-6. doi: 10.1073/pnas.1213515110. Epub 2012 Dec 17.

Abstract

Hepatitis C virus (HCV) replication is dependent on microRNA 122 (miR-122), a liver-specific microRNA that recruits Argonaute 2 to the 5' end of the viral genome, stabilizing it and slowing its decay both in cell-free reactions and in infected cells. Here we describe the RNA degradation pathways against which miR-122 provides protection. Transfected HCV RNA is degraded by both the 5' exonuclease Xrn1 and 3' exonuclease exosome complex, whereas replicating RNA within infected cells is degraded primarily by Xrn1 with no contribution from the exosome. Consistent with this, sequencing of the 5' and 3' ends of RNA degradation intermediates in infected cells confirmed that 5' decay is the primary pathway for HCV RNA degradation. Xrn1 knockdown enhances HCV replication, indicating that Xrn1 decay and the viral replicase compete to set RNA abundance within infected cells. Xrn1 knockdown and miR-122 supplementation have equal, redundant, and nonadditive effects on the rate of viral RNA decay, indicating that miR-122 protects HCV RNA from 5' decay. Nevertheless, Xrn1 knockdown does not rescue replication of a viral mutant defective in miR-122 binding, indicating that miR-122 has additional yet uncharacterized function(s) in the viral life cycle.

摘要

丙型肝炎病毒(HCV)的复制依赖于 microRNA 122(miR-122),miR-122 是一种肝脏特异性 microRNA,它将 Argonaute 2 募集到病毒基因组的 5'端,稳定病毒并减缓其在无细胞反应和感染细胞中的降解。在这里,我们描述了 miR-122 提供保护的 RNA 降解途径。转染的 HCV RNA 可被 5'外切核酸酶 Xrn1 和 3'外切核酸酶 exosome 复合物降解,而感染细胞内复制的 RNA 主要被 Xrn1 降解,exosome 没有贡献。与此一致,对感染细胞中 RNA 降解中间产物的 5'和 3'末端进行测序证实,5'降解是 HCV RNA 降解的主要途径。Xrn1 敲低增强了 HCV 的复制,表明 Xrn1 降解和病毒复制酶竞争在感染细胞内设定 RNA 丰度。Xrn1 敲低和 miR-122 补充对病毒 RNA 降解的速率具有相同、冗余和非加性的影响,表明 miR-122 保护 HCV RNA 免受 5'降解。然而,Xrn1 敲低不能挽救 miR-122 结合缺陷的病毒突变体的复制,表明 miR-122 在病毒生命周期中具有额外的、尚未确定的功能。

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