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一个位于大麦黄矮病毒(PAV)基因组5'端4.5至5千碱基处的序列强烈刺激无帽mRNA的翻译。

A sequence located 4.5 to 5 kilobases from the 5' end of the barley yellow dwarf virus (PAV) genome strongly stimulates translation of uncapped mRNA.

作者信息

Wang S, Miller W A

机构信息

Plant Pathology Department, Iowa State University, Ames 50011-1020, USA.

出版信息

J Biol Chem. 1995 Jun 2;270(22):13446-52. doi: 10.1074/jbc.270.22.13446.

Abstract

An infectious, in vitro transcript from a full-length cDNA clone of the barley yellow dwarf virus (PAV serotype) genome translated efficiently in a wheat germ translation extract. Deletions in a region that we call the 3' translational enhancer, located between bases 4,513 and 5,009 in the 5,677-base genome, reduced translation of the 5'-proximal open reading frames from uncapped RNA by at least 30-fold. Deletions elsewhere in all but the 5' end of the genome had no effect on translation. Presence of a m7G(5')pp(5')G cap on the 5' end fully restored translational efficiency of transcripts lacking the 3' translational enhancer. The translation enhancer reduced inhibition of translation by free cap analog, did not affect RNA stability, and did not function in reticulocyte lysates. When placed in the 3'-untranslated region of uncapped mRNA encoding the beta-glucuronidase gene, the translation enhancer stimulated translation more than 80-fold, in the presence of the viral, but not a plasmid-derived, 5' leader. A polyadenylate tail could not substitute for the 3' translation enhancer. These observations provide an extreme example, in terms of distance from the 5' end and level of stimulation, of an mRNA in which a sequence near the 3' end stimulates translation.

摘要

大麦黄矮病毒(PAV血清型)基因组全长cDNA克隆的感染性体外转录本在小麦胚翻译提取物中能有效翻译。在我们称为3'翻译增强子的区域(位于5677个碱基的基因组中第4513至5009个碱基之间)进行缺失,会使来自无帽RNA的5'近端开放阅读框的翻译减少至少30倍。除基因组5'端外其他位置的缺失对翻译没有影响。5'端存在m7G(5')pp(5')G帽可完全恢复缺乏3'翻译增强子的转录本的翻译效率。该翻译增强子可降低游离帽类似物对翻译的抑制作用,不影响RNA稳定性,且在网织红细胞裂解物中不起作用。当置于编码β-葡萄糖醛酸酶基因的无帽mRNA的3'非翻译区时,在病毒来源而非质粒来源的5'前导序列存在的情况下,该翻译增强子可使翻译增强80多倍。聚腺苷酸尾巴不能替代3'翻译增强子。就与5'端的距离和刺激水平而言,这些观察结果提供了一个极端的例子,即3'端附近的序列刺激翻译的mRNA。

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