cis-Acting signals that promote genome replication in rotavirus mRNA.
cis-Acting signals that promote genome replication in rotavirus mRNA.
复制标题
促进轮状病毒 mRNA 基因组复制的顺式作用信号。
DOI：
10.1128/jvi.70.6.3961-3971.1996
复制
发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Ramig,RF
中科院分区：
文献类型：
--
作者：
Patton,JT;Wentz,M;Xiaobo,J;Ramig,RF
先前的研究已经表明轮状病毒核心具有相关的复制酶活性，其可以指导在无细胞系统中从病毒mRNA合成双链RNA（D. Y. Chen C.，马缨丹属问：Y. Zeng，M. J. Wentz，M. Gorziglia，M. K. Estes和R. F. Ramig，J. Virol. 68：7030-7039，1994）。为了确定轮状病毒mRNA中对RNA复制重要的顺式作用信号，产生了含有内部和末端缺失的基因8转录物和将基因8特异性3 '末端序列连接到非病毒序列末端的嵌合转录物。在无细胞系统中对这些RNA的分析导致在基因8 mRNA中鉴定出对RNA复制必需的顺式作用信号和两种顺式作用信号，这两种顺式作用信号虽然对复制不是必需的，但用于增强该过程。基本复制信号的序列位于基因8 mRNA的3'末端，并且由于其高度保守的性质，可能是所有11种病毒mRNA的共同特征。通过基因8 mRNA的位点特异性突变，发现3'必需信号的位置-1，-2，-5，-6和-7的残基对于促进RNA复制特别重要。在无细胞体系中，增强复制的顺式作用信号之一位于基因8 mRNA的3'非翻译区（UTR）的5'端附近，而另一个则位于信使的5'非翻译区。5' UTR中增强信号的存在提高了轮状病毒mRNA的5'和3'末端在基因组复制期间可以彼此相互作用和/或与病毒复制酶相互作用的可能性。
A previous study has shown that rotavirus cores have an associated replicase activity which can direct the synthesis of double-stranded RNA from viral mRNA in a cell-free system (D. Y. Chen, C. Q.-Y. Zeng, M. J. Wentz, M. Gorziglia, M. K. Estes, and R. F. Ramig, J. Virol. 68:7030-7039, 1994). To define the cis-acting signals in rotavirus mRNA that are important for RNA replication, gene 8 transcripts which contained internal and terminal deletions and chimeric transcripts which linked gene 8-specific 3'-terminal sequences to the ends of nonviral sequences were generated. Analysis of these RNAs in the cell-free system led to the identification of a cis-acting signal in the gene 8 mRNA which is essential for RNA replication and two cis-acting signals which, while not essential for replication, serve to enhance the process. The sequence of the essential replication signal is located at the extreme 3' end of the gene 8 mRNA and, because of its highly conserved nature, is probably a common feature of all 11 viral mRNAs. By site-specific mutagenesis of the gene 8 mRNA, residues at positions -1, -2, -5, -6, and -7 of the 3' essential signal were found to be particularly important for promoting RNA replication. One of the cis-acting signals shown to enhance the replication in the cell-free system was located near the 5' end of the 3' untranslated region (UTR) of the gene 8 mRNA, while remarkably the other was located in the 5' UTR of the message. The existence of an enhancement signal in the 5' UTR raises the possibility that the 5' and 3' ends of the rotavirus mRNA may interact with each other and/or with the viral replicase during genome replication.