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冠状病毒5'端基因组区域顺式作用RNA元件的结构与功能保守性

Structural and functional conservation of cis-acting RNA elements in coronavirus 5'-terminal genome regions.

作者信息

Madhugiri Ramakanth, Karl Nadja, Petersen Daniel, Lamkiewicz Kevin, Fricke Markus, Wend Ulrike, Scheuer Robina, Marz Manja, Ziebuhr John

机构信息

Institute of Medical Virology, Justus Liebig University, Giessen, Germany.

Faculty of Mathematics and Computer Science, Friedrich Schiller University, Jena, Germany; European Virus Bioinformatics Center, Jena, Germany.

出版信息

Virology. 2018 Apr;517:44-55. doi: 10.1016/j.virol.2017.11.025. Epub 2017 Dec 6.

DOI:10.1016/j.virol.2017.11.025
PMID:29223446
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7112051/
Abstract

Structure predictions suggest a partial conservation of RNA structure elements in coronavirus terminal genome regions. Here, we determined the structures of stem-loops (SL) 1 and 2 of two alphacoronaviruses, human coronavirus (HCoV) 229E and NL63, by RNA structure probing and studied the functional relevance of these putative cis-acting elements. HCoV-229E SL1 and SL2 mutants generated by reverse genetics were used to study the effects on viral replication of single-nucleotide substitutions predicted to destabilize the SL1 and SL2 structures. The data provide conclusive evidence for the critical role of SL1 and SL2 in HCoV-229E replication and, in some cases, revealed parallels with previously characterized betacoronavirus SL1 and SL2 elements. Also, we were able to rescue viable HCoV-229E mutants carrying replacements of SL2 with equivalent betacoronavirus structural elements. The data obtained in this study reveal a remarkable degree of structural and functional conservation of 5'-terminal RNA structural elements across coronavirus genus boundaries.

摘要

结构预测表明冠状病毒末端基因组区域的RNA结构元件存在部分保守性。在此，我们通过RNA结构探测确定了两种甲型冠状病毒，即人冠状病毒（HCoV）229E和NL63的茎环（SL）1和2的结构，并研究了这些假定的顺式作用元件的功能相关性。利用反向遗传学产生的HCoV-229E SL1和SL2突变体来研究预测会破坏SL1和SL2结构的单核苷酸取代对病毒复制的影响。这些数据为SL1和SL2在HCoV-229E复制中的关键作用提供了确凿证据，并且在某些情况下，揭示了与先前表征的乙型冠状病毒SL1和SL2元件的相似之处。此外，我们能够拯救携带用等效乙型冠状病毒结构元件替换SL2的有活力的HCoV-229E突变体。本研究获得的数据揭示了跨冠状病毒属边界的5'末端RNA结构元件在结构和功能上的显著保守程度。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7e7b962cf94c/gr8_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/d7c15a9a8112/gr1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/c2b25bfb3632/gr2_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/80df7c3736c7/gr3_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7e930df9d342/gr4_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7b1236f69133/gr5_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/e22b8d9c2ca1/gr6_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/bf3956ccc208/gr7_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7e7b962cf94c/gr8_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/d7c15a9a8112/gr1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/c2b25bfb3632/gr2_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/80df7c3736c7/gr3_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7e930df9d342/gr4_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7b1236f69133/gr5_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/e22b8d9c2ca1/gr6_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/bf3956ccc208/gr7_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/345f/7112051/7e7b962cf94c/gr8_lrg.jpg

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