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花椰菜花叶病毒逆转录酶并非通过酿酒酵母中的核糖体移码机制产生。

The cauliflower mosaic virus reverse transcriptase is not produced by the mechanism of ribosomal frameshifting in Saccharomyces cerevisiae.

作者信息

Wurch T, Guidasci T, Geldreich A, Lebeurier G, Mesnard J M

机构信息

Institut de Biologie Moléculaire des Plantes du CNRS, Université Louis Pasteur, Strasbourg, France.

出版信息

Virology. 1991 Feb;180(2):837-41. doi: 10.1016/0042-6822(91)90103-i.

Abstract

The capsid protein and the reverse transcriptase of cauliflower mosaic virus (CaMV) are encoded by two genes (ORF IV and ORF V) that lie in different translation reading frames. A comparison can be drawn between the synthesis of both CaMV proteins and the fusion protein in a yeast retrotransposon, Ty, resulting from a +1 frameshifting event which fuses two out-of-phase ORFs encoding the structural protein and the reverse transcriptase of Ty. For this reason, we constructed a yeast expression vector containing CaMV ORF VII fused to CaMV ORF III by a fragment of 452 bp including the overlapping region of ORF IV and ORF V, ORF VII and ORF III being used as reporter genes. We characterized two proteins (22 and 50 kDa) synthesized from this plasmid in the yeast expression system. We demonstrated that the 50-kDa polypeptide is not synthesized from a +1 frameshifting event but is probably a dimeric form of the 22-kDa protein. From this result we conclude that the CaMV reverse transcriptase is not produced by a mechanism of ribosomal frameshifting.

摘要

花椰菜花叶病毒(CaMV)的衣壳蛋白和逆转录酶由位于不同翻译阅读框的两个基因(ORF IV和ORF V)编码。可以对CaMV这两种蛋白的合成与酵母逆转座子Ty中的融合蛋白的合成进行比较,Ty中的融合蛋白是由一个+1移码事件产生的,该事件使两个编码Ty结构蛋白和逆转录酶的异相开放阅读框融合。因此,我们构建了一个酵母表达载体,其中CaMV ORF VII通过一段452 bp的片段与CaMV ORF III融合,该片段包括ORF IV和ORF V的重叠区域,ORF VII和ORF III用作报告基因。我们对在酵母表达系统中从该质粒合成的两种蛋白(22 kDa和50 kDa)进行了表征。我们证明50 kDa的多肽不是由+1移码事件合成的,而可能是22 kDa蛋白的二聚体形式。根据这一结果,我们得出结论,CaMV逆转录酶不是通过核糖体移码机制产生的。

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