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异源烟草花叶病毒运动蛋白和嵌合运动蛋白基因对细胞间和长距离运动的影响。

Influence of heterologous tobamovirus movement protein and chimeric-movement protein genes on cell-to-cell and long-distance movement.

作者信息

Deom C M, He X Z, Beachy R N, Weissinger A K

机构信息

Department of Plant Pathology, University of Georgia, Athens 30602-7274.

出版信息

Virology. 1994 Nov 15;205(1):198-209. doi: 10.1006/viro.1994.1635.

DOI:10.1006/viro.1994.1635
PMID:7975216
Abstract

Sunn-hemp mosaic tobamovirus (SHMV) moves slowly from cell to cell in Nicotiana tabacum cv. Xanthi, but fails to move long distance. To determine the role of the SHMV movement protein (MP) in cell-to-cell and long-distance movement in tobacco, the SHMV MP gene was inserted into a TMV-cDNA clone that had approximately the 5'-half of the endogenous MP gene deleted. RNA transcripts inoculated onto tobacco induced systemic infections by 8 days postinoculation. Sequence analysis of the MP genes from purified virus isolated from systemically infected leaf tissue indicated that chimeric SHMV/TMV MP genes had been generated through RNA-RNA recombination within the 3'-termini of the MP gene sequences. When exchanged for the MP gene of TMV, three of four chimeric MP genes analyzed provided long-distance movement function for the hybrid viruses in tobacco. Two of the three hybrid viruses that moved long distance showed enhanced cell-to-cell movement relative to a recombinant TMV that expressed the intact SHMV MP gene. These observations suggest that the C-terminus of the TMV MP contains a determinant that can influence cell-to-cell movement in tobacco. A recombinant virus, TLSM, that expressed the intact SHMV MP gene exhibited cell-to-cell movement that was intermediate to SHMV and TMV, but failed to produce coat protein and was defective in long-distance movement. To further examine the role of the SHMV MP gene in long-distance movement, transgenic N. tabacum cv. Xanthi that expressed the wild-type SHMV MP gene were generated and found to facilitate rapid and efficient long-distance movement of a TMV mutant that contained a dysfunctional MP gene. Therefore, the inability of SHMV to systemically infect tobacco is a function of virus components and sequences other than those encoded by the SHMV MP gene.

摘要

太阳麻花叶烟草花叶病毒(SHMV)在烟草品种黄花烟草中从一个细胞缓慢移动到另一个细胞,但无法进行长距离移动。为了确定SHMV移动蛋白(MP)在烟草细胞间和长距离移动中的作用,将SHMV MP基因插入到一个TMV-cDNA克隆中,该克隆的内源性MP基因的大约5'端已被删除。接种到烟草上的RNA转录本在接种后8天诱导了系统感染。对从系统感染的叶片组织中分离出的纯化病毒的MP基因进行序列分析表明,嵌合的SHMV/TMV MP基因是通过MP基因序列3'端内的RNA-RNA重组产生的。当用四个嵌合MP基因中的三个替换TMV的MP基因时,分析发现它们为烟草中的杂种病毒提供了长距离移动功能。在三个能进行长距离移动的杂种病毒中,有两个相对于表达完整SHMV MP基因的重组TMV显示出增强的细胞间移动能力。这些观察结果表明,TMV MP的C末端含有一个可以影响烟草细胞间移动的决定因素。一种表达完整SHMV MP基因的重组病毒TLSM,其细胞间移动能力介于SHMV和TMV之间,但不能产生外壳蛋白,并且在长距离移动方面存在缺陷。为了进一步研究SHMV MP基因在长距离移动中的作用,构建了表达野生型SHMV MP基因的转基因烟草品种黄花烟草,发现它促进了一个含有功能失调MP基因的TMV突变体的快速高效长距离移动。因此,SHMV无法系统感染烟草是由病毒成分和序列决定的,而非SHMV MP基因所编码的那些成分和序列。

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