A PSEUDOKNOT-LIKE STRUCTURE REQUIRED FOR EFFICIENT SELF-CLEAVAGE OF HEPATITIS DELTA-VIRUS RNA
A PSEUDOKNOT-LIKE STRUCTURE REQUIRED FOR EFFICIENT SELF-CLEAVAGE OF HEPATITIS DELTA-VIRUS RNA
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DOI：
10.1038/350434a0
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发表时间：
1991-04-04
期刊：
影响因子：
64.8
通讯作者：
BEEN, MD
中科院分区：
文献类型：
--
作者：
PERROTTA, AT;BEEN, MD
丁型肝炎病毒基因组和反基因组RNA含有自切割位点，假设其在复制过程中加工病毒RNA 1-3。 自切割仅需要二价阳离子1-3，并且在基因组位点由少于85个核苷酸的序列介导4。 我们认为基因组自切割序列元件4和来自反基因组RNA的相应序列可以产生相关的二级结构。 反基因组序列的区域，从提出的结构预测，合成和显示是足够的自我切割。 通过反基因组序列的位点特异性诱变获得了形成三级相互作用的两个茎的证据。 在10 M甲酰胺或5 M尿素中的有效自切割，也是基因组序列5的性质，依赖于两个茎中的碱基配对。 但在不存在变性剂的情况下，不需要切割位点远端的茎，这表明三级相互作用稳定了自切割所需的结构。
HEPATITIS delta virus genomic and antigenomic RNAs contain a self-cleavage site hypothesized to function in processing the viral RNA during replication 1-3. Self-cleavage requires only a divalent cation 1-3 and is mediated at the genomic site by a sequence of less than 85 nucleotides 4. We propose that the genomic self-cleaving sequence element 4 and a corresponding sequence from the antigenomic RNA could generate related secondary structures. The region of the antigenomic sequence, predicted from the proposed structure, was synthesized and shown to be sufficient for self-cleavage. Evidence for two stems which form a tertiary interaction was obtained by site-specific mutagenesis of the antigenomic sequence. Efficient self-cleavage in 10 M formamide or 5 M urea, also a property of the genomic sequence 5, was dependent on base-pairing in both stems. But in the absence of denaturants, the stem distal to the site of cleavage was not required, suggesting that the tertiary interaction stabilizes the structure required for self-cleavage.