Interaction of the host protein NbDnaJ with Potato virus X minus-strand stem-loop 1 RNA and capsid protein affects viral replication and movement
Interaction of the host protein NbDnaJ with Potato virus X minus-strand stem-loop 1 RNA and capsid protein affects viral replication and movement
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DOI：
10.1016/j.bbrc.2011.11.137
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发表时间：
2012-01-06
影响因子：
3.1
通讯作者：
Kim, Kook-Hyung
中科院分区：
文献类型：
--
作者：
Cho, Sang-Yun;Cho, Won Kyong;Kim, Kook-Hyung
植物病毒必须与宿主细胞成分相互作用才能复制并在细胞间移动。在马铃薯X病毒（PVX）的情况下，它携带病毒复制和移动所必需的茎环1（SL 1）RNA。使用二维电泳西北印迹分析，我们以前确定了几个主机的蛋白质结合到SL 1 RNA。其中，我们进一步表征了来自本氏烟草（Nicotiana benthamiana）的命名为NbDnaJ的DnaJ样蛋白。电泳迁移率变动分析证实，NbDnaJ仅结合到SL 1负链RNA，和双分子荧光互补（BiFC）表明，NbDnaJ与PVX衣壳蛋白（CP）相互作用。使用一系列的缺失突变体，NbDnaJ的C-末端区域被发现是必不可少的与PVX CP的相互作用。NbDnaJ的表达显着变化后，感染不同的植物病毒，如PVX，烟草花叶病毒，黄瓜花叶病毒，但不同的病毒种类。在瞬时实验中，PVX的复制和运动都被抑制在植物中，过度表达NbDnaJ，但加速在植物中，NbDnaJ是沉默的。总之，我们认为新鉴定的NbDnaJ通过与SL 1（-）RNA和PVX CP相互作用在PVX复制和运动中起作用。(C)2011 Elsevier Inc. All rights reserved.
Plant viruses must interact with host cellular components to replicate and move from cell to cell. In the case of Potato virus X (PVX), it carries stem-loop 1 (SL1) RNA essential for viral replication and movement. Using two-dimensional electrophoresis northwestern blot analysis, we previously identified several host proteins that bind to SL1 RNA. Of those, we further characterized a DnaJ-like protein from Nicotiana benthamiana named NbDnaJ. An electrophoretic mobility shift assay confirmed that NbDnaJ binds only to SL1 minus-strand RNA, and bimolecular fluorescence complementation (BiFC) indicated that NbDnaJ interacts with PVX capsid protein (CP). Using a series of deletion mutants, the C-terminal region of NbDnaJ was found to be essential for the interaction with PVX CP. The expression of NbDnaJ significantly changed upon infection with different plant viruses such as PVX, Tobacco mosaic virus, and Cucumber mosaic virus, but varied depending on the viral species. In transient experiments, both PVX replication and movement were inhibited in plants that over-expressed NbDnaJ but accelerated in plants in which NbDnaJ was silenced. In summary, we suggest that the newly identified NbDnaJ plays a role in PVX replication and movement by interacting with SL1(-) RNA and PVX CP. (C) 2011 Elsevier Inc. All rights reserved.