Specificity of Plant Rhabdovirus Cell-to-Cell Movement
Specificity of Plant Rhabdovirus Cell-to-Cell Movement
复制标题
植物弹状病毒细胞间运动的特异性
DOI：
10.1128/jvi.00296-19
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发表时间：
2019-05
影响因子：
5.4
通讯作者：
Li Zhenghe
中科院分区：
文献类型：
--
作者：
Zhou Xin;Lin Wenye;Sun Kai;Wang Shuo;Zhou Xueping;Jackson Andrew O.;Li Zhenghe
植物弹状病毒的局部运输可能涉及病毒核衣壳通过MP门控胞间连丝，但分子机制尚未完全了解。我们已经进行了互补试验与MP编码的五个不同的弹状病毒，以评估其运动的特异性。每个弹状病毒MP补充了两个正链RNA病毒的MP缺陷突变体的运动，这两个正链RNA病毒具有不同的运动策略。与此形成鲜明对比的是，两种重组植物弹状病毒的细胞间运动是高度特异性的，需要它们的同源MP。我们已经表明，这些弹状病毒MP是本地化的细胞周边和与细胞膜，它们只与它们的同源核衣壳核心蛋白相互作用。这些相互作用能够将病毒核衣壳核心蛋白从其复制位点重定向到细胞周边。我们的研究为植物弹状病毒的特异性胞间和胞内运输提供了一个模型，该模型可能适用于其他负链RNA病毒。正链RNA病毒运动蛋白（MPs）通常缺乏序列特异性核酸结合活性，并与相关和不相关病毒显示跨家族运动互补性。负链RNA植物弹状病毒编码的MP与其他植物病毒对应物的结构和功能相关性有限，但细胞间运输的确切机制尚不清楚。在这项研究中，我们首先分析了MPs编码的五种不同的弹状病毒支持细胞间运动的两个正链RNA病毒通过使用反式互补试验的能力。五个弹状病毒MP中的每一个都补充了番茄花叶病毒和马铃薯X病毒的MP缺陷突变体的运动。相比之下，苦菜黄网核弹状病毒（SYNV）和番茄黄斑驳相关细胞弹状病毒（TYMaV）的重组MP缺失突变体的运动仅由其相应的MP拯救，即，SYNV sc 4和TYMaV P3。亚细胞分离分析表明，SYNV sc 4和TYMaV P3与细胞膜外周相关。分裂泛素膜酵母双杂交试验证明了膜相关弹状病毒MP仅与其同源核蛋白（N）和磷蛋白（P）的特异性相互作用。更重要的是，SYNV sc 4-N和sc 4-P相互作用将一定比例的N-P复合物从核复制位点引导到细胞外周的点状位点，该位点与胞间连丝部分共定位。我们的数据表明，植物弹状病毒的细胞到细胞的运动是高度特异性的，并建议同源MP-核衣壳核心蛋白的相互作用所需的细胞内和细胞间的贩运。重要性植物弹状病毒的局部运输可能涉及病毒核衣壳通过MP门控胞间连丝，但分子机制尚未完全了解。我们已经进行了互补试验与MP编码的五个不同的弹状病毒，以评估其运动的特异性。每个弹状病毒MP补充了两个正链RNA病毒的MP缺陷突变体的运动，这两个正链RNA病毒具有不同的运动策略。与此形成鲜明对比的是，两种重组植物弹状病毒的细胞间运动是高度特异性的，需要它们的同源MP。我们已经表明，这些弹状病毒MP是本地化的细胞周边和与细胞膜，它们只与它们的同源核衣壳核心蛋白相互作用。这些相互作用能够将病毒核衣壳核心蛋白从其复制位点重定向到细胞周边。我们的研究为植物弹状病毒的特异性胞间和胞内运输提供了一个模型，该模型可能适用于其他负链RNA病毒。
Local transport of plant rhabdoviruses likely involves the passage of viral nucleocapsids through MP-gated plasmodesmata, but the molecular mechanisms are not fully understood. We have conducted complementation assays with MPs encoded by five distinct rhabdoviruses to assess their movement specificity. Each of the rhabdovirus MPs complemented the movement of MP-defective mutants of two positive-stranded RNA viruses that have different movement strategies. In marked contrast, cell-to-cell movement of two recombinant plant rhabdoviruses was highly specific in requiring their cognate MPs. We have shown that these rhabdovirus MPs are localized to the cell periphery and associate with cellular membranes, and that they interact only with their cognate nucleocapsid core proteins. These interactions are able to redirect viral nucleocapsid core proteins from their sites of replication to the cell periphery. Our study provides a model for the specific inter- and intracellular trafficking of plant rhabdoviruses that may be applicable to other negative-stranded RNA viruses. ABSTRACT Positive-stranded RNA virus movement proteins (MPs) generally lack sequence-specific nucleic acid-binding activities and display cross-family movement complementarity with related and unrelated viruses. Negative-stranded RNA plant rhabdoviruses encode MPs with limited structural and functional relatedness with other plant virus counterparts, but the precise mechanisms of intercellular transport are obscure. In this study, we first analyzed the abilities of MPs encoded by five distinct rhabdoviruses to support cell-to-cell movement of two positive-stranded RNA viruses by using trans-complementation assays. Each of the five rhabdovirus MPs complemented the movement of MP-defective mutants of tomato mosaic virus and potato X virus. In contrast, movement of recombinant MP deletion mutants of sonchus yellow net nucleorhabdovirus (SYNV) and tomato yellow mottle-associated cytorhabdovirus (TYMaV) was rescued only by their corresponding MPs, i.e., SYNV sc4 and TYMaV P3. Subcellular fractionation analyses revealed that SYNV sc4 and TYMaV P3 were peripherally associated with cell membranes. A split-ubiquitin membrane yeast two-hybrid assay demonstrated specific interactions of the membrane-associated rhabdovirus MPs only with their cognate nucleoproteins (N) and phosphoproteins (P). More importantly, SYNV sc4-N and sc4-P interactions directed a proportion of the N-P complexes from nuclear sites of replication to punctate loci at the cell periphery that partially colocalized with the plasmodesmata. Our data show that cell-to-cell movement of plant rhabdoviruses is highly specific and suggest that cognate MP-nucleocapsid core protein interactions are required for intra- and intercellular trafficking. IMPORTANCE Local transport of plant rhabdoviruses likely involves the passage of viral nucleocapsids through MP-gated plasmodesmata, but the molecular mechanisms are not fully understood. We have conducted complementation assays with MPs encoded by five distinct rhabdoviruses to assess their movement specificity. Each of the rhabdovirus MPs complemented the movement of MP-defective mutants of two positive-stranded RNA viruses that have different movement strategies. In marked contrast, cell-to-cell movement of two recombinant plant rhabdoviruses was highly specific in requiring their cognate MPs. We have shown that these rhabdovirus MPs are localized to the cell periphery and associate with cellular membranes, and that they interact only with their cognate nucleocapsid core proteins. These interactions are able to redirect viral nucleocapsid core proteins from their sites of replication to the cell periphery. Our study provides a model for the specific inter- and intracellular trafficking of plant rhabdoviruses that may be applicable to other negative-stranded RNA viruses.
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