RNA PSEUDOKNOT DOMAIN OF TOBACCO MOSAIC-VIRUS CAN FUNCTIONALLY SUBSTITUTE FOR A POLY(A) TAIL IN PLANT AND ANIMAL-CELLS
RNA PSEUDOKNOT DOMAIN OF TOBACCO MOSAIC-VIRUS CAN FUNCTIONALLY SUBSTITUTE FOR A POLY(A) TAIL IN PLANT AND ANIMAL-CELLS
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DOI：
10.1101/gad.4.7.1149
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发表时间：
1990-07-01
影响因子：
10.5
通讯作者：
WALBOT, V
中科院分区：
文献类型：
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作者：
GALLIE, DR;WALBOT, V
许多RNA病毒的基因组终止于类似于tRNA的L-构象的三级结构，并且该结构被人tRNA特异性酶如氨酰-tRNA合成酶识别。烟草花叶病毒（TMV）RNA的几乎整个3“-非翻译区（UTR）参与延伸的三级结构，除了tRNA样结构之外，还包含位于紧邻上游的假结结构域。虽然这些结构的功能还不清楚，但它们对病毒至关重要。我们证明，与非腺苷酸化的mRNA相比，将204个碱基的TMV 3“-非翻译区添加到外源mRNA构建体中可以使基因表达增加高达100倍。TMV的3 ''-UTR等于或大于一个多聚腺苷酸化的尾部，在电穿孔的双子叶植物和单子叶植物原生质体中增强基因表达。TMV 3“-UTR在功能上类似于多腺苷酸化尾，因为它增加mRNA稳定性和翻译，并且必须位于3”末端以有效发挥功能。在中国仓鼠卵巢细胞中观察到对表达的类似影响，表明该序列在广泛的真核生物中起作用。当扩展的三级结构被解剖，上游假结域被发现是主要负责增加表达。然而，包含tRNA样结构对于完全调节是重要的。
The genomes of many RNA viruses terminate in a tertiary structure similar to the L-conformation of tRNAs and this structure is recognized by man tRNA-specific enzymes such as aminoacyl-tRNA synthetase. Virtually the entire 3''-untranslated region (UTR) of tobacco mosaic virus (TMV) RNA is involved in an extended tertiary structure containing, in addition to a tRNA-like structure, a pseudoknot domain that lies immediately upstream. Although the functions of these structures are not well understood, they are essential to the virus. We demonstrate that the addition of the 204-base TMV 3''-untranslated region to foreign mRNA constructs can increase gene expression up to 100-fold compared to nonadenylated mRNA. The 3''-UTR of TMV was equal to or greater than a polyadenylated tail in enhancing gene expression in electroporated dicot and monocot protoplasts. The TMV 3''-UTR is functionally similar to a polyadenylated tail in that it increases mRNA stability and translation and must be positioned at the 3'' terminus to function efficiently. Similar effects on expression were observed in Chinese hamster ovary cells, demonstrating that the sequence functions in a wide range of eukaryotes. When the extended tertiary structure was dissected, the upstream pseudoknot domain was found to be largely responsible for increasing expression. The inclusion of the tRNA-like structure, however, was important for full regulation.