RNA dimerization plays a role in ribosomal frameshifting of the SARS coronavirus.
RNA dimerization plays a role in ribosomal frameshifting of the SARS coronavirus.
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DOI：
10.1093/nar/gks1361
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发表时间：
2013-02-01
影响因子：
14.9
通讯作者：
Hennig M
中科院分区：
文献类型：
--
作者：
Ishimaru D;Plant EP;Sims AC;Yount BL Jr;Roth BM;Eldho NV;Pérez-Alvarado GC;Armbruster DW;Baric RS;Dinman JD;Taylor DR;Hennig M
参与程序化-1核糖体移框（-1 PRF）的信使RNA编码信号是典型的双茎发夹(H)型假结（pks）。我们之前描述了严重急性呼吸综合征（SARS）冠状病毒（CoV）刺激-1 PRF的不寻常的三茎假结。第三个茎环的保守存在暗示了一个重要的迄今未知的功能。在这里，我们提出了描述SARS假结第三茎的结构和功能的新信息。我们通过嵌入在SARS-CoV Stem 3中的回文序列发现了RNA二聚化。进一步的体外分析表明，SARS-CoV RNA二聚体通过“亲吻”环环相互作用组装。我们还表明，在生理温度和镁的存在下，环环接吻复合物的形成变得更有效。当回文序列发生突变时，体外RNA二聚化被消除，帧移从15%降低到5.7%。此外，由SARS-CoV的Stem 3密码子沉默改变引起的无法二聚体化改变了病毒生长动力学，并影响了感染细胞中基因组和亚基因组RNA的水平。这些结果表明，由SARS假结形成的同二聚体RNA复合物发生在细胞环境中，涉及Stem 3调节-1 PRF的环环亲吻相互作用在亚基因组和全长RNA合成中发挥作用。
Messenger RNA encoded signals that are involved in programmed -1 ribosomal frameshifting (-1 PRF) are typically two-stemmed hairpin (H)-type pseudoknots (pks). We previously described an unusual three-stemmed pseudoknot from the severe acute respiratory syndrome (SARS) coronavirus (CoV) that stimulated -1 PRF. The conserved existence of a third stem–loop suggested an important hitherto unknown function. Here we present new information describing structure and function of the third stem of the SARS pseudoknot. We uncovered RNA dimerization through a palindromic sequence embedded in the SARS-CoV Stem 3. Further in vitro analysis revealed that SARS-CoV RNA dimers assemble through ‘kissing’ loop–loop interactions. We also show that loop–loop kissing complex formation becomes more efficient at physiological temperature and in the presence of magnesium. When the palindromic sequence was mutated, in vitro RNA dimerization was abolished, and frameshifting was reduced from 15 to 5.7%. Furthermore, the inability to dimerize caused by the silent codon change in Stem 3 of SARS-CoV changed the viral growth kinetics and affected the levels of genomic and subgenomic RNA in infected cells. These results suggest that the homodimeric RNA complex formed by the SARS pseudoknot occurs in the cellular environment and that loop–loop kissing interactions involving Stem 3 modulate -1 PRF and play a role in subgenomic and full-length RNA synthesis.
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影响因子：
5.6
作者：
Brierley I;Rolley NJ;Jenner AJ;Inglis SC
通讯作者：
Inglis SC
DOI：
10.1073/pnas.0905046106
发表时间：
2009-08-04
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1073/pnas.0608668104
发表时间：
2007-04-03
影响因子：
11.1
作者：
Hansen, Thomas M.;Reihani, S. Nader S.;Sorensen, Michael A.
通讯作者：
Sorensen, Michael A.
影响因子：
5.8
作者：
Larkin, M. A.;Blackshields, G.;Higgins, D. G.
通讯作者：
Higgins, D. G.