Structural and functional analysis of dengue virus RNA.
Structural and functional analysis of dengue virus RNA.
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DOI：
10.1002/0470058005.ch9
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发表时间：
2006-01-01
影响因子：
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通讯作者：
Gamarnik, Andrea V
中科院分区：
文献类型：
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作者：
Alvarez, Diego E;Lodeiro, Maria F;Gamarnik, Andrea V
存在于RNA病毒的5'和3' UTR的序列和结构在翻译、RNA合成和病毒组装的起始和调节中起关键作用。在登革病毒以及其他蚊媒黄病毒中，基因组末端互补序列的存在介导长距离RNA-RNA相互作用。登革病毒RNA显示基因组环化和病毒活力所需的两对互补序列（CS和UAR）。为了研究这些RNA-RNA相互作用参与病毒生命周期的分子机制，我们开发了登革病毒复制子系统。蚊子和哺乳动物细胞中复制子的RNA转染允许区分参与翻译和RNA合成的RNA元件。我们发现，在RNA的5'或3'末端的CS或UAR内的突变干扰碱基配对，并不显著影响输入RNA的翻译，但严重损害或破坏RNA合成。此外，UAR序列的系统突变分析表明，除了在RNA环化中的作用外，UAR内的特定核苷酸对于有效的RNA合成也很重要。
Sequences and structures present at the 5' and 3' UTRs of RNA viruses play crucial roles in the initiation and regulation of translation, RNA synthesis and viral assembly. In dengue virus, as well as in other mosquito-borne flaviviruses, the presence of complementary sequences at the ends of the genome mediate long-range RNA-RNA interactions. Dengue virus RNA displays two pairs of complementary sequences (CS and UAR) required for genome circularization and viral viability. In order to study the molecular mechanism by which these RNA-RNA interactions participate in the viral life cycle, we developed a dengue virus replicon system. RNA transfection of the replicon in mosquito and mammalian cells allows discrimination between RNA elements involved in translation and RNA synthesis. We found that mutations within CS or UAR at the 5' or 3' ends of the RNA that interfere with base pairing did not significantly affect translation of the input RNA but seriously compromised or abolished RNA synthesis. Furthermore, a systematic mutational analysis of UAR sequences indicated that, beside the role in RNA cyclization, specific nucleotides within UAR are also important for efficient RNA synthesis.