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玄参花叶病毒(花椰菜花叶病毒组)的基因VI在全长RNA转录本上基因的转录后表达中发挥作用。

Gene VI of figwort mosaic virus (caulimovirus group) functions in posttranscriptional expression of genes on the full-length RNA transcript.

作者信息

Gowda S, Wu F C, Scholthof H B, Shepherd R J

机构信息

Department of Plant Pathology, University of Kentucky, Lexington 40546.

出版信息

Proc Natl Acad Sci U S A. 1989 Dec;86(23):9203-7. doi: 10.1073/pnas.86.23.9203.

Abstract

Experimental evidence for a molecular function for gene VI of the caulimoviruses is presented. Based on experiments with the figwort mosaic virus (FMV), it appears that gene VI has a role in the posttranscriptional expression of the closely packed genes (VII and I-V), which appear on the larger, full-length RNA transcript of this virus. Gene VI with its flanking 5'/3' expression signals included as a separate plasmid during electroporation of DNA into protoplasts of Nicotiana edwardsonii shows an unusual type of transactivation of a chloramphenicol acetyltransferase (CAT) gene fused at its 5' end to a small open reading frame (gene VII) of the long 5' leader of the full-length RNA transcript of the FMV genome. The level of activity of the CAT gene is increased up to 20-fold over the activity of control plasmids when gene VI is included in the electroporation mixture. Mutagenesis of the coding portions of gene VI of pGS1 RVI, a transactivating plasmid used in the electroporation experiments, demonstrated that it was probably the polypeptide product of gene VI that was responsible for the transactivating effect. Experiments with various portions of the 5' leader of the large, full-length RNA of FMV showed that the coding region of gene VII is necessary for the transactivation event. Clones of cauliflower mosaic virus (CaMV) or FMV with intact gene VI were found to reciprocally transactivate gene VII-CAT fusions (FMV) or gene I-CAT fusions (CaMV) located downstream of the 5' leader sequences of either viral genome.

摘要

本文提供了花椰菜花叶病毒基因VI分子功能的实验证据。基于对玄参花叶病毒(FMV)的实验,基因VI似乎在紧密排列的基因(VII和I-V)的转录后表达中发挥作用,这些基因出现在该病毒的较大全长RNA转录本上。在将DNA电穿孔导入爱德华氏烟草原生质体的过程中,包含侧翼5'/3'表达信号的基因VI作为一个单独的质粒,显示出对氯霉素乙酰转移酶(CAT)基因的一种不寻常的反式激活作用,该CAT基因在其5'端与FMV基因组全长RNA转录本长5'前导序列的一个小开放阅读框(基因VII)融合。当基因VI包含在电穿孔混合物中时,CAT基因的活性水平比对照质粒的活性提高了20倍。在电穿孔实验中使用的反式激活质粒pGS1 RVI的基因VI编码部分的诱变表明,可能是基因VI的多肽产物负责反式激活作用。对FMV大的全长RNA的5'前导序列不同部分的实验表明,基因VII的编码区域对于反式激活事件是必需的。发现具有完整基因VI的花椰菜花叶病毒(CaMV)或FMV克隆可相互反式激活位于任一病毒基因组5'前导序列下游的基因VII-CAT融合体(FMV)或基因I-CAT融合体(CaMV)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8830/298462/f85274cafd58/pnas00290-0163-a.jpg

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