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20S RNA纳罗病毒在酿酒酵母中对抗SKI1/XRN1的抗病毒活性。

20S RNA narnavirus defies the antiviral activity of SKI1/XRN1 in Saccharomyces cerevisiae.

作者信息

Esteban Rosa, Vega Lorena, Fujimura Tsutomu

机构信息

Instituto de Microbiología Bioquímica, Departamento de Microbiología y Genética, Consejo Superior de Investigaciones Científicas/Universidad de Salamanca, Salamanca 37007, Spain.

出版信息

J Biol Chem. 2008 Sep 19;283(38):25812-20. doi: 10.1074/jbc.M804400200. Epub 2008 Jul 18.

Abstract

20S RNA virus is a persistent positive strand RNA virus found in Saccharomyces cerevisiae. We previously observed that the virus generated in vivo from a launching vector possessed the correct RNA termini without extra sequences. Here we present evidence that the SKI1/XRN1 5'-exonuclease plays a major role in the elimination of the non-viral upstream sequences from the primary transcripts. The virus, once generated, however, is fairly unaffected by overexpression or deletion of SKI1/XRN1. By contrast, the copy number of the L-A double-stranded RNA virus in the same host is greatly increased by the deletion of SKI1/XRN1, and overexpression of the gene cured L-A virus from the cells at a high frequency. 20S RNA virus, unlike L-A virus, has a strong secondary structure at its 5'-end: the first four nucleotides are G, and they are buried at the bottom of a long stem structure, features known to inhibit the SKI1/XRN1 5'-exonuclease progression. Mutations that weakened the 5'-stem structure made 20S RNA virus vulnerable to SKI1/XRN1. These results, together with the data on L-A virus, indicate a strong anti-RNA virus activity of SKI1/XRN1. Given that 20S RNA virus resides and replicates in the cytoplasm without a protective capsid, our results suggest that the strong secondary structure at the 5'-end is crucial for the 20S RNA virus to evade the host SKI1/XRN1 defense.

摘要

20S RNA病毒是在酿酒酵母中发现的一种持久性正链RNA病毒。我们之前观察到,由启动载体在体内产生的病毒具有正确的RNA末端,没有额外的序列。在此我们提供证据表明,SKI1/XRN1 5'-外切核酸酶在从初级转录本中消除非病毒上游序列方面起主要作用。然而,一旦病毒产生,它相当不受SKI1/XRN1过表达或缺失的影响。相比之下,同一宿主中L-A双链RNA病毒的拷贝数因SKI1/XRN1的缺失而大大增加,并且该基因的过表达能高频地从细胞中清除L-A病毒。与L-A病毒不同,20S RNA病毒在其5'-末端具有很强的二级结构:前四个核苷酸是G,它们埋藏在一个长茎结构的底部,这些特征已知会抑制SKI1/XRN1 5'-外切核酸酶的作用进程。削弱5'-茎结构的突变使20S RNA病毒易受SKI1/XRN1的影响。这些结果与关于L-A病毒的数据一起,表明SKI1/XRN1具有很强的抗RNA病毒活性。鉴于20S RNA病毒在没有保护性衣壳的情况下存在于细胞质中并进行复制,我们的结果表明,5'-末端的强二级结构对于20S RNA病毒逃避宿主SKI1/XRN1防御至关重要。

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