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番茄黄化曲叶病毒(TYLCV)衣壳蛋白在植物和昆虫细胞中的核输入

Nuclear import of the capsid protein of tomato yellow leaf curl virus (TYLCV) in plant and insect cells.

作者信息

Kunik T, Palanichelvam K, Czosnek H, Citovsky V, Gafni Y

机构信息

Department of Genetics, Agricultural Research Organization, Bet Dagan, Israel.

出版信息

Plant J. 1998 Feb;13(3):393-9. doi: 10.1046/j.1365-313x.1998.00037.x.

Abstract

The tomato yellow leaf curl virus (TYLCV) found in Israel is a whitefly-transmitted monopartite geminivirus. Although geminiviruses have been found in the nuclei of phloem-associated cells, the mechanism of viral invasion is poorly understood. The possible role of the TYLCV capsid protein (CP), the only known component of the viral coat, in virus transport into the host cell nucleus was investigated by monitoring its specific nuclear accumulation in plant and insect cells. CP was fused to the beta-glucuronidase (GUS) reporter enzyme to assay nuclear import in petunia protoplasts, and micro-injection of purified fluorescently labeled CP was used to examine its nuclear uptake in Drosophila embryos. Both assays demonstrated that TYLCV CP is transported into plant- and insect-cell nuclei by an active process of nuclear import via a nuclear localization signal (NLS)-specific pathway. Using the GUS assay and deletion analysis, the TYLCV CP NLS sequence was identified in the amino-terminus of the protein.

摘要

在以色列发现的番茄黄化曲叶病毒(TYLCV)是一种由烟粉虱传播的单分体双生病毒。尽管在韧皮部相关细胞的细胞核中发现了双生病毒,但病毒入侵的机制仍知之甚少。通过监测其在植物和昆虫细胞中的特异性核积累,研究了TYLCV衣壳蛋白(CP)(病毒衣壳唯一已知的成分)在病毒转运到宿主细胞核中的可能作用。将CP与β-葡萄糖醛酸酶(GUS)报告酶融合,以检测矮牵牛原生质体中的核输入,并使用显微注射纯化的荧光标记CP来检查其在果蝇胚胎中的核摄取。两种检测方法均表明,TYLCV CP通过核定位信号(NLS)特异性途径的主动核输入过程转运到植物和昆虫细胞核中。使用GUS检测和缺失分析,在该蛋白的氨基末端鉴定出了TYLCV CP NLS序列。

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