SHAPE analysis of the RNA secondary structure of the Mouse Hepatitis Virus 5' untranslated region and N-terminal nsp1 coding sequences.
SHAPE analysis of the RNA secondary structure of the Mouse Hepatitis Virus 5' untranslated region and N-terminal nsp1 coding sequences.
复制标题
DOI：
10.1016/j.virol.2014.11.001
复制
发表时间：
2015-01-15
期刊：
影响因子：
3.7
通讯作者：
Leibowitz, Julian L.
中科院分区：
文献类型：
--
作者：
Yang, Dong;Liu, Pinghua;Wudeck, Elyse V.;Giedroc, David P.;Leibowitz, Julian L.
关键词：
用Shape技术分析了MHV-A59基因组5‘端最大的474个核苷酸的RNA二级结构，其中包含了缺陷干扰RNA复制所需的最小的5’顺式作用区域。所生成的结构与SL1至SL4以及最近预测的两个二级结构元件S5和SL5A的特征一致。形状为之前没有在MHV中进行功能研究的另外四个茎环提供了生化支持。尽管序列差异较大，但MHV-A59、BCOV和SARS-CoV的5‘端结构预测是相似的。病毒基因组内RNA、病毒外基因组RNA和体外合成的RNA的形状反应模式相似，表明N蛋白或其他蛋白质与病毒RNA的结合不能保护RNA免受脂透性SHAPE试剂的影响。反向遗传实验表明，NSP1编码序列中的SL5C和SL6不是病毒复制所必需的。对MHV完整的5‘顺式作用区域的RNA二级结构进行了形状分析。β冠状病毒的RNA二级结构从5端到SL5都是保守的。SL5C和SL6不是病毒复制所必需的。
SHAPE technology was used to analyze RNA secondary structure of the 5′ most 474 nts of the MHV-A59 genome encompassing the minimal 5′ cis-acting region required for defective interfering RNA replication. The structures generated were in agreement with previous characterizations of SL1 through SL4 and two recently predicted secondary structure elements, S5 and SL5A. SHAPE provided biochemical support for four additional stem–loops not previously functionally investigated in MHV. Secondary structure predictions for 5′ regions of MHV-A59, BCoV and SARS-CoV were similar despite high sequence divergence. The pattern of SHAPE reactivity of in virio genomic RNA, ex virio genomic RNA, and in vitro synthesized RNA was similar, suggesting that binding of N protein or other proteins to virion RNA fails to protect the RNA from reaction with lipid permeable SHAPE reagent. Reverse genetic experiments suggested that SL5C and SL6 within the nsp1 coding sequence are not required for viral replication. SHAPE analysis of RNA secondary structure was performed on the complete 5′ cis-acting region of MHV. The RNA secondary structures of betacoronaviruses are conserved from the 5 end through SL5. SL5C and SL6 are not required for viral replication.
登录
查看更多内容
影响因子：
3.7
作者：
Brockway SM;Denison MR
通讯作者：
Denison MR
影响因子：
5
作者：
Gorbalenya AE;Enjuanes L;Ziebuhr J;Snijder EJ
通讯作者：
Snijder EJ
影响因子：
5.4
作者：
Kang, Hyojeung;Feng, Min;Leibowitz, Julian L.
通讯作者：
Leibowitz, Julian L.
影响因子：
5.6
作者：
Laing, Christian;Schlick, Tamar
通讯作者：
Schlick, Tamar
影响因子：
3.7
作者：
Chen, Shih-Cheng;Olsthoorn, Rene C. L.
通讯作者：
Olsthoorn, Rene C. L.