Woolaway K E, Lazaridis K, Belsham G J, Carter M J, Roberts L O
School of Biomedical and Life Sciences, University of Surrey, Guildford GU2 7XH, United Kingdom.
J Virol. 2001 Nov;75(21):10244-9. doi: 10.1128/JVI.75.21.10244-10249.2001.
Rhopalosiphum padi virus (RhPV) is one of several picorna-like viruses that infect insects; sequence analysis has revealed distinct differences between these agents and mammalian picornaviruses. RhPV has a single-stranded positive-sense RNA genome of about 10 kb; unlike the genomes of Picornaviridae, however, this genome contains two long open reading frames (ORFs). ORF1 encodes the virus nonstructural proteins, while the downstream ORF, ORF2, specifies the structural proteins. Both ORFs are preceded by long untranslated regions (UTRs). The intergenic UTR is known to contain an internal ribosome entry site (IRES) which directs non-AUG-initiated translation of ORF2. We have examined the 5' UTR of RhPV for IRES activity by translating synthetic dicistronic mRNAs containing this sequence in a variety of systems. We now report that the 5' UTR contains an element which directs internal initiation of protein synthesis from an AUG codon in mammalian, plant, and Drosophila in vitro translation systems. In contrast, the encephalomyocarditis virus IRES functions only in the mammalian system. The RhPV 5' IRES element has features in common with picornavirus IRES elements, in that no coding sequence is required for IRES function, but also with cellular IRES elements, as deletion analysis indicates that this IRES element does not have sharply defined boundaries.
禾谷缢管蚜病毒(RhPV)是几种感染昆虫的类微小核糖核酸病毒之一;序列分析揭示了这些病原体与哺乳动物微小核糖核酸病毒之间存在明显差异。RhPV具有一个约10 kb的单链正义RNA基因组;然而,与微小核糖核酸病毒科的基因组不同,该基因组包含两个长开放阅读框(ORF)。ORF1编码病毒非结构蛋白,而下游的ORF2则指定结构蛋白。两个ORF之前都有长的非翻译区(UTR)。已知基因间UTR包含一个内部核糖体进入位点(IRES),它指导ORF2的非AUG起始翻译。我们通过在多种系统中翻译包含该序列的合成双顺反子mRNA,研究了RhPV的5'UTR的IRES活性。我们现在报告,5'UTR包含一个元件,它在哺乳动物、植物和果蝇体外翻译系统中指导从AUG密码子开始的蛋白质合成内部起始。相比之下,脑心肌炎病毒IRES仅在哺乳动物系统中起作用。RhPV 5'IRES元件与微小核糖核酸病毒IRES元件有共同特征,即IRES功能不需要编码序列,但也与细胞IRES元件有共同特征,因为缺失分析表明该IRES元件没有明确界定的边界。