Deletion of the s2m RNA Structure in the Avian Coronavirus Infectious Bronchitis Virus and Human Astrovirus Results in Sequence Insertions.
Deletion of the s2m RNA Structure in the Avian Coronavirus Infectious Bronchitis Virus and Human Astrovirus Results in Sequence Insertions.
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DOI：
10.1128/jvi.00038-23
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发表时间：
2023-03-30
影响因子：
5.4
通讯作者：
Bickerton E
中科院分区：
文献类型：
--
作者：
Keep S;Dowgier G;Lulla V;Britton P;Oade M;Freimanis G;Tennakoon C;Jonassen CM;Tengs T;Bickerton E
冠状病毒感染多种宿主物种，导致人类和动物出现一系列疾病。冠状病毒基因组由一个含有许多RNA结构的大型正义单链RNA分子组成。其中一个结构为s2m，由41个核苷酸组成，位于3 ‘非翻译区（3 ’ UTR）内，在一些冠状病毒物种（包括传染性支气管炎病毒（IBV）、严重急性呼吸综合征冠状病毒（SARS-CoV）和SARS-CoV-2）以及其他病原体（包括人类星状病毒）之间共享。利用反向遗传系统生成重组病毒，我们研究了IBV复制对s2m结构的要求。IBV是一种全球分布的具有重要经济意义的γ冠状病毒，感染家禽引起呼吸道疾病。三个核苷酸的缺失被预测会破坏s2m的典型结构，或者与s2m对应的核苷酸的缺失会影响病毒在体外的复制。对缺失s2m序列的重组IBV进行体外传代，在缺失的地方插入了36个核苷酸，被鉴定为由侧翼序列的重复组成。在人类星状病毒序列中删除s2m序列后，也观察到类似的结果。RNA建模表明，与s2m对应的核苷酸的缺失影响了IBV 3 ' UTR中存在的其他RNA结构。我们的研究结果表明，IBV和人类星状病毒都倾向于占据与s2m对应的基因组位置的核苷酸，这与特定的s2m序列无关。冠状病毒感染许多物种，包括人类和动物，对牲畜，特别是家禽产生重大影响。冠状病毒RNA基因组由参与病毒复制的结构元件组成，人们对这些元件的作用知之甚少。本文研究了禽γ冠状病毒传染性支气管炎病毒（一种具有重要经济意义的病毒病原体）在复制过程中对RNA结构元件s2m的需求。利用反向遗传学技术产生具有中断或缺失的s2m的重组ibv，我们发现s2m在细胞培养中不是病毒复制所必需的；然而，气管组织的复制减少，表明s2m在自然宿主中的作用。这些病毒以及缺乏s2m序列的人类星状病毒的传代显示出对核苷酸占据的偏好，独立于s2m序列。RNA建模表明，s2m的缺失可能会对其他基本RNA结构产生负面影响。
Coronaviruses infect a wide variety of host species, resulting in a range of diseases in both humans and animals. The coronavirus genome consists of a large positive-sense single-stranded molecule of RNA containing many RNA structures. One structure, denoted s2m and consisting of 41 nucleotides, is located within the 3′ untranslated region (3′ UTR) and is shared between some coronavirus species, including infectious bronchitis virus (IBV), severe acute respiratory syndrome coronavirus (SARS-CoV), and SARS-CoV-2, as well as other pathogens, including human astrovirus. Using a reverse genetic system to generate recombinant viruses, we investigated the requirement of the s2m structure in the replication of IBV, a globally distributed economically important Gammacoronavirus that infects poultry causing respiratory disease. Deletion of three nucleotides predicted to destabilize the canonical structure of the s2m or the deletion of the nucleotides corresponding to s2m impacted viral replication in vitro. In vitro passaging of the recombinant IBV with the s2m sequence deleted resulted in a 36-nucleotide insertion in place of the deletion, which was identified to be composed of a duplication of flanking sequences. A similar result was observed following serial passage of human astrovirus with a deleted s2m sequence. RNA modeling indicated that deletion of the nucleotides corresponding to the s2m impacted other RNA structures present in the IBV 3′ UTR. Our results indicated for both IBV and human astrovirus a preference for nucleotide occupation in the genome location corresponding to the s2m, which is independent of the specific s2m sequence. IMPORTANCE Coronaviruses infect many species, including humans and animals, with substantial effects on livestock, particularly with respect to poultry. The coronavirus RNA genome consists of structural elements involved in viral replication whose roles are poorly understood. We investigated the requirement of the RNA structural element s2m in the replication of the Gammacoronavirus infectious bronchitis virus, an economically important viral pathogen of poultry. Using reverse genetics to generate recombinant IBVs with either a disrupted or deleted s2m, we showed that the s2m is not required for viral replication in cell culture; however, replication is decreased in tracheal tissue, suggesting a role for the s2m in the natural host. Passaging of these viruses as well as human astrovirus lacking the s2m sequence demonstrated a preference for nucleotide occupation, independent of the s2m sequence. RNA modeling suggested deletion of the s2m may negatively impact other essential RNA structures.
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影响因子：
3.7
作者：
Armesto M;Cavanagh D;Britton P
通讯作者：
Britton P
影响因子：
3.5
作者：
Ferretti, Luca;Tennakoon, Chandana;Ribeca, Paolo
通讯作者：
Ribeca, Paolo
影响因子：
2.8
作者：
Cook, JKA;Orbell, SJ;Huggins, MB
通讯作者：
Huggins, MB
DOI：
10.1007/978-1-59745-181-9_3
发表时间：
2008
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Jonassen CM
通讯作者：
Jonassen CM
影响因子：
64.5
作者：
Brierley I;Digard P;Inglis SC
通讯作者：
Inglis SC