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Molecular analyzes of the mechanism of plant virus pathogenicity

植物病毒致病机制的分子分析

基本信息

批准号：
08306003
负责人：
FURUSAWA Iwao
金额：
$ 15.87万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08306003/
关键词：
CaMV TMV CMV PVX ToMV RNA polymerase host factor host specificity RDV RNA複製酵素植物ウイルスゲノム解析病原性遺伝子ポリメラーゼ全身感染

项目摘要

We have comprehensively studied the mechanism of genome replication of various plant viruses. Chronological analyzes of the accumulation of viral gene products in cauliflower mosaic virus (CaMV)-infected turnip protoplasts showed that the expression of each gene is differentially regulated. It has also been suggested that subgenomic RNAs as well as the polycistronic 35S RNA are involved in the expression of CaMV genes, indicating that CaMV uses a unique strategy for the gene expression during virus multiplication. Two domains of the 180K protein gene of tobacco mosaic virus (TMV)-OM,each containing the sequence motifs of methyltransferase or RNA polymerase, have shown to exhibit the binding activity with TMV-RNA.By immunoaffinity chromatography using antibodies specfic to the domains, heterodimers consisting of 130K and 180K viral proteins, which exhibited TMV-RNA dependent RNA synthesis in vitro, were isolated from TMV-infected tobacco leaves. We have analyzed Arabidopsis thaliana mutants designated cum1 and cum2, in which the multiplication of cucumber mosaic virsu (CMV) is delayd. Furthermore, we have been trying to clone the CUM1 gene by the positional cloning procedure. We have isolated solubilized RNA polymerase synthesizing subgenomic RNA from tobacco protoplasts inoculated with potato virus V (PVX). Amino acid 979 of the 130-and 180-kDa proteins of tomato mosaic tobamovirus (ToMV) plays an important role in overcoming the tomato Tm-1 resistance gene. Introduction of a single amino acid substitution (Gln979 to Ile) into the ToMV proteins rendered the mutant virtually unable to replicate in tomato cells while it replicated perfectly in tobacco cells, indicating the importance of the region surrounding the amino acid 979 in terms of host specificity determinants.

我们全面研究了多种植物病毒的基因组复制机制。对花椰菜花叶病毒（CaMV）感染的萝卜原生质体中病毒基因产物积累的时间分析表明，每个基因的表达受到差异性调节。也有人提出亚基因组RNA以及多顺反子35S RNA参与CaMV基因的表达，表明CaMV在病毒增殖过程中使用独特的基因表达策略。烟草花叶病毒（TMV）-OM的180K蛋白基因的两个结构域均含有甲基转移酶或RNA聚合酶的序列基序，已显示出与TMV-RNA的结合活性。通过使用该结构域特异性抗体的免疫亲和层析，得到由130K和180K病毒蛋白组成的异二聚体，其表现出从 TMV 感染的烟叶中分离出 TMV-RNA 依赖性 RNA 体外合成。我们分析了拟南芥突变体 cum1 和 cum2，其中黄瓜花叶病毒 (CMV) 的增殖被延迟。此外，我们一直在尝试通过定位克隆方法克隆CUM1基因。我们从接种马铃薯病毒 V (PVX) 的烟草原生质体中分离出合成亚基因组 RNA 的溶解性 RNA 聚合酶。番茄花叶病毒 (ToMV) 130 kDa 和 180 kDa 蛋白的第 979 位氨基酸在克服番茄 Tm-1 抗性基因方面发挥着重要作用。在ToMV蛋白中引入单个氨基酸取代（Gln979到Ile）使得突变体几乎无法在番茄细胞中复制，而它在烟草细胞中完美复制，这表明氨基酸979周围的区域在宿主特异性决定因素方面的重要性。