Role of Rice Stripe Virus NSvc4 in Cell-to-Cell Movement and Symptom Development in Nicotiana benthamiana.
Role of Rice Stripe Virus NSvc4 in Cell-to-Cell Movement and Symptom Development in Nicotiana benthamiana.
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DOI：
10.3389/fpls.2012.00269
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发表时间：
2012
影响因子：
5.6
通讯作者：
Zhou X
中科院分区：
文献类型：
--
作者：
Xu Y;Zhou X
我们之前的工作已经证明，水稻条纹病毒（RSV）的NSvc4蛋白是一种细胞间运动蛋白。然而，RSV通过胞间连丝（plasmodesmata， PD）传播的机制尚不清楚。在这里，我们提供的证据表明，NSvc4在肌动蛋白丝和内质网网络上移动，而不是微管，到达细胞壁PD。使用不同的抑制剂破坏细胞骨架，改变NSvc4对PD的定位，从而阻碍本菌烟对RSV的感染。序列分析和缺失诱变实验表明，NSvc4的n端125个氨基酸（AAs）决定PD的靶向性，横跨AAs 106-125的跨膜结构域对PD的定位至关重要。我们还发现NSvc4蛋白可以定位到感染细胞的叶绿体上。对缺失突变体的分析表明，n端73 AAs对叶绿体定位至关重要。此外，从马铃薯病毒X （PVX）载体中表达的NSvc4在系统感染的benthamiana叶片中比PVX单独表达更严重的疾病症状。NSvc4在夜蛾9细胞中的表达不会引起小管的形成，而是在质膜处形成点状灶。这些发现为我们理解RSV感染宿主植物的运动机制提供了新的线索。
Our previous work has demonstrated that the NSvc4 protein of Rice stripe virus (RSV) functions as a cell-to-cell movement protein. However, the mechanisms whereby RSV traffics through plasmodesmata (PD) are unknown. Here we provide evidence that the NSvc4 moves on the actin filament and endoplasmic reticulum network, but not microtubules, to reach cell wall PD. Disruption of cytoskeleton using different inhibitors altered NSvc4 localization to PD, thus impeding RSV infection of Nicotiana benthamiana. Sequence analyses and deletion mutagenesis experiment revealed that the N-terminal 125 amino acids (AAs) of the NSvc4 determine PD targeting and that a transmembrane domain spanning AAs 106–125 is critical for PD localization. We also found that the NSvc4 protein can localize to chloroplasts in infected cells. Analyses using deletion mutants revealed that the N-terminal 73 AAs are essential for chloroplast localization. Furthermore, expression of NSvc4 from a Potato virus X (PVX) vector resulted in more severe disease symptoms than PVX alone in systemically infected N. benthamiana leaves. Expression of NSvc4 in Spodoptera frugiperda 9 cells did not elicit tubule formation, but instead resulted in punctate foci at the plasma membrane. These findings shed new light on our understanding of the movement mechanisms whereby RSV infects host plants.
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