Stone D M, Almond J W, Brangwyn J K, Belsham G J
Department of Microbiology, University of Reading, Whiteknights, United Kingdom.
J Virol. 1993 Oct;67(10):6215-23. doi: 10.1128/JVI.67.10.6215-6223.1993.
Poliovirus (PV) RNA is translated by a cap-independent mechanism involving the internal entry of ribosomes onto the 5' noncoding region (NCR). Using the vaccinia virus-T7 RNA polymerase transient expression system, we showed previously that deletion of certain individual predicted secondary structures within the PV 5' NCR rendered the element defective in directing internal initiation when assayed alone. However, these defective 5' NCRs were functional when coexpressed within cells with full-length PV cDNA (N. Percy, G. J. Belsham, J. K. Brangwyn, M. Sullivan, D. M. Stone, and J. W. Almond, J. Virol. 66:1695-1701, 1992). We have extended the study to demonstrate that when these predicted secondary structures are deleted in combination, the enhanced activity in the presence of the full-length PV cDNA is still observed. Indeed, a poliovirus 5' NCR devoid of all predicted secondary structures is capable of initiating protein synthesis under these conditions. Surprisingly, we also found that this enhancement of activity requires neither any PV protein nor the inhibition of cap-dependent translation. The results indicate that the defective PV 5' NCR elements can be complemented in trans by functional 5' NCRs in a highly sequence specific manner.
脊髓灰质炎病毒(PV)RNA通过一种不依赖帽子结构的机制进行翻译,该机制涉及核糖体在5'非编码区(NCR)的内部进入。利用痘苗病毒-T7 RNA聚合酶瞬时表达系统,我们之前表明,单独检测时,PV 5' NCR内某些预测的二级结构的缺失使该元件在指导内部起始方面存在缺陷。然而,当这些有缺陷的5' NCR与全长PV cDNA在细胞内共表达时,它们是有功能的(N. Percy、G. J. Belsham、J. K. Brangwyn、M. Sullivan、D. M. Stone和J. W. Almond,《病毒学杂志》66:1695-1701,1992)。我们扩展了这项研究,以证明当这些预测的二级结构组合缺失时,在全长PV cDNA存在的情况下,仍能观察到增强的活性。事实上,在这些条件下,一个没有所有预测二级结构的脊髓灰质炎病毒5' NCR能够起始蛋白质合成。令人惊讶的是,我们还发现这种活性增强既不需要任何PV蛋白,也不需要抑制依赖帽子结构的翻译。结果表明,有缺陷的PV 5' NCR元件可以被功能性5' NCR以高度序列特异性的方式反式互补。