The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
复制标题
DOI：
10.1371/journal.ppat.1010589
复制
发表时间：
2022-06
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
作者：
病毒 RNA (vRNA) 基因组的非编码区对于基因表达的调控至关重要。特别是，假结 (PK) 结构存在于多种 RNA 分子中，具有多种作用。口蹄疫病毒 (FMDV) vRNA 的 5' 非翻译区 (5' UTR) 比小核糖核酸病毒家族的其他病毒长得多，并且由许多独特的结构基序组成，其中包括多个（2、3 或 4 个，具体取决于病毒株）串联连接的推定 PK。 PK 在 FMDV 感染中的作用尚不完全清楚。在这里，我们利用生物信息学、亚基因组复制子和重组病毒研究了 FMDV 生命周期中 PK 的结构保守性和重要性。我们的结果表明，尽管所有 FMDV 中都保留了两个或多个 PK，但缺乏 PK 的复制子仍具有复制能力，尽管水平较低。此外，在竞争实验中，具有少于两个（0 或 1）个 PK 结构的 GFP FMDV 复制子被具有 4 个 PK 的 mCherry FMDV wt 复制子竞争，而具有 2 或 4 个 PK 的 GFP 复制子则不然。额外的 PK 提供的这种明显的复制优势与 FMDV 野外分离株基因组中至少两个 PK 的维持相关。尽管复制子缺乏任何保留复制能力的 PK，但完全缺乏 PK 的病毒无法存活，并且至少有一个 PK 对于感染病毒的恢复至关重要，这表明 PK 在病毒体组装中发挥作用。因此，我们的研究指出了 PK 在 vRNA 复制和病毒粒子组装中的作用，从而加深了对 FMDV 复制的分子生物学以及 PK 在 RNA 功能中更广泛作用的理解。口蹄疫（FMD）是一种高度传染性的病毒性疾病，对全球畜牧业构成持续威胁，因为它在世界许多地区仍然流行。开发新的口蹄疫控制措施可能在全球范围内具有重大的经济和社会价值，但这需要更深入地了解口蹄疫病毒复制的分子机制。人们对病毒 RNA 基因组的许多特征仍然知之甚少，包括一系列假定的假结 (PK) 结构的作用，预计 RNA 会在其中折叠起来。在这项研究中，我们证实了 FMDV 基因组中这些 PK 的保守性和结构，并表明至少有一个 PK 的存在对于感染性病毒颗粒的产生至关重要。然而，两个或多个 PK 的存在在复制竞争实验中是有利的。这些结果与所有已知 FMDV 野外分离株的基因组至少包含两个 PK 结构的观察结果相关。我们建议 PK 在复制和病毒粒子组装中的作用。
Non-coding regions of viral RNA (vRNA) genomes are critically important in the regulation of gene expression. In particular, pseudoknot (PK) structures, which are present in a wide range of RNA molecules, have a variety of roles. The 5′ untranslated region (5′ UTR) of foot-and-mouth disease virus (FMDV) vRNA is considerably longer than in other viruses from the picornavirus family and consists of a number of distinctive structural motifs that includes multiple (2, 3 or 4 depending on the virus strain) putative PKs linked in tandem. The role(s) of the PKs in the FMDV infection are not fully understood. Here, using bioinformatics, sub-genomic replicons and recombinant viruses we have investigated the structural conservation and importance of the PKs in the FMDV lifecycle. Our results show that despite the conservation of two or more PKs across all FMDVs, a replicon lacking PKs was replication competent, albeit at reduced levels. Furthermore, in competition experiments, GFP FMDV replicons with less than two (0 or 1) PK structures were outcompeted by a mCherry FMDV wt replicon that had 4 PKs, whereas GFP replicons with 2 or 4 PKs were not. This apparent replicative advantage offered by the additional PKs correlates with the maintenance of at least two PKs in the genomes of FMDV field isolates. Despite a replicon lacking any PKs retaining the ability to replicate, viruses completely lacking PK were not viable and at least one PK was essential for recovery of infections virus, suggesting a role for the PKs in virion assembly. Thus, our study points to roles for the PKs in both vRNA replication and virion assembly, thereby improving understanding the molecular biology of FMDV replication and the wider roles of PK in RNA functions. Foot-and-mouth disease (FMD) is a highly contagious viral disease posing a constant threat to the global livestock industry, as it remains endemic in many parts of the world. The development of novel control measures for FMD could have major economic and social value worldwide, however, this will require a deeper understanding of the molecular mechanisms of FMDV replication. A number of features of the viral RNA genome remain poorly understood, including the role of a series of putative pseudoknot (PK) structures, where the RNA is predicted to fold back on itself. In this study, we confirmed the conservation and structure of these PKs in the FMDV genome and showed that the presence of at least one is essential for production of infectious virus particles. However, the presence of two or more PKs was advantageous in replication competition experiments. These results correlate with the observation that genomes of all known FMDV field isolates include at least two PK structures. We suggest roles for the PKs in both replication and virion assembly.
登录
查看更多内容
影响因子：
5.4
作者：
Herod MR;Ferrer-Orta C;Loundras EA;Ward JC;Verdaguer N;Rowlands DJ;Stonehouse NJ
通讯作者：
Stonehouse NJ
DOI：
10.1093/bioinformatics/btp250
发表时间：
2009-08-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Darty K;Denise A;Ponty Y
通讯作者：
Ponty Y
影响因子：
6.7
作者：
Herod MR;Gold S;Lasecka-Dykes L;Wright C;Ward JC;McLean TC;Forrest S;Jackson T;Tuthill TJ;Rowlands DJ;Stonehouse NJ
通讯作者：
Stonehouse NJ
影响因子：
4.5
作者：
Karabiber, Fethullah;McGinnis, Jennifer L.;Weeks, Kevin M.
通讯作者：
Weeks, Kevin M.
DOI：
10.1073/pnas.1219988110
发表时间：
2013-04-02
影响因子：
11.1
作者：
Hajdin, Christine E.;Bellaousov, Stanislav;Weeks, Kevin M.
通讯作者：
Weeks, Kevin M.