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Residues R192 and K225 in RNA-Binding Pocket of Tobacco Vein Banding Mosaic Virus CP Control Virus Cell-to-Cell Movement and Replication
Molecular Plant-Microbe Interactions ( IF 3.5 ) Pub Date : 2021-07-21 , DOI: 10.1094/mpmi-09-20-0265-r
Zhi-Yong Yan 1 , Xiao-Jie Xu 1 , Le Fang 1 , De-Jie Cheng 1 , Yan-Ping Tian 1 , Chao Geng 1 , Xiang-Dong Li 1 , Jari P. T. Valkonen 2
Affiliation  

Potyviruses move to neighboring cells in the form of virus particles or a coat protein (CP)-containing ribonucleoprotein complex. However, the precise roles of RNA-binding residues in potyviral CP in viral cell-to-cell movement remain to be elucidated. In this study, we predicted the three-dimensional model of tobacco vein banding mosaic virus (TVBMV)-encoded CP and found nine residues presumably located in the CP RNA-binding pocket. Substitutions of the two basic residues at positions 192 and 225 (R192 and K225) with either alanine, cysteine, or glutamic acid abolished TVBMV cell-to-cell and systemic movement in Nicotiana benthamiana plants. These substitutions also reduced the replication of the mutant viruses. Results from the electrophoretic mobility shift assay showed that the RNA-binding activity of mutant CPs derived from R192 or K225 substitutions was significantly lower than that of wild-type CP. Analysis of purified virus particles showed that mutant viruses with R192 or K225 substitutions formed RNA-free virus-like particles. Mutations of R192 and K225 did not change the CP plasmodesmata localization. The wild-type TVBMV CP could rescue the deficient cell-to-cell movement of mutant viruses. Moreover, deletion of any of the other seven residues also abolished TVBMV cell-to-cell movement and reduced the CP RNA-binding activity. The corresponding nine residues in watermelon mosaic virus CP were also found to play essential roles in virus cell-to-cell movement. In conclusion, residues R192 and K225 in the CP RNA-binding pocket are critical for viral RNA binding and affect both virus replication and cell-to-cell movement.

Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.



中文翻译:

烟草静脉带状花叶病毒 CP 的 RNA 结合袋中的 R192 和 K225 残留 控制病毒细胞间运动和复制

Potyvirus 以病毒颗粒或含有外壳蛋白 (CP) 的核糖核蛋白复合物的形式移动到邻近细胞。然而,Potyviral CP 中 RNA 结合残基在病毒细胞间运动中的确切作用仍有待阐明。在这项研究中,我们预测了烟草静脉带状花叶病毒 (TVBMV) 编码的 CP 的三维模型,并发现了 9 个可能位于 CP RNA 结合口袋中的残基。用丙氨酸、半胱氨酸或谷氨酸取代第 192 和 225 位(R 192和 K 225)的两个碱性残基,消除了本氏烟草中TVBMV 细胞间和全身运动植物。这些替换也减少了突变病毒的复制。电泳迁移率变化分析的结果表明,源自 R 192或 K 225取代的突变 CP 的 RNA 结合活性显着低于野生型 CP。对纯化病毒颗粒的分析表明,带有 R 192或 K 225取代的突变病毒形成了不含 RNA 的病毒样颗粒。R 192和 K 225 的突变没有改变 CP 胞间连丝定位。野生型 TVBMV CP 可以挽救突变病毒的细胞间运动缺陷。此外,删除其他七个残基中的任何一个也会消除 TVBMV 细胞间运动并降低 CP RNA 结合活性。还发现西瓜花叶病毒 CP 中相应的 9 个残基在病毒细胞间运动中起重要作用。总之,CP RNA 结合口袋中的残基 R 192和 K 225对病毒 RNA 结合至关重要,并影响病毒复制和细胞间运动。

版权所有 © 2021 作者。这是在 CC BY-NC-ND 4.0 国际许可下分发的开放获取文章。

更新日期:2021-07-21
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