p24(G1) Encoded by Grapevine Leafroll-Associated Virus 1 Suppresses RNA Silencing and Elicits Hypersensitive Response-Like Necrosis in Nicotiana Species.
p24(G1) Encoded by Grapevine Leafroll-Associated Virus 1 Suppresses RNA Silencing and Elicits Hypersensitive Response-Like Necrosis in Nicotiana Species.
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葡萄藤相关病毒1编码的p24（G1）抑制RNA沉默并引发烟草属物种的过敏反应样坏死
DOI：
10.3390/v12101111
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发表时间：
2020-09-30
期刊：
影响因子：
--
通讯作者：
Cheng YQ
中科院分区：
文献类型：
--
作者：
Zhang CW;Liu Q;Zeng Q;Huang WT;Wang Q;Cheng YQ
葡萄叶卷相关病毒1 （GLRaV-1）是葡萄叶卷病的主要病原。然而，glrav1与植物细胞相互作用的分子机制尚不清楚。通过农杆菌浸润介导的RNA沉默实验，我们发现glrav1 p24蛋白（p24G1）可作为RNA沉默抑制因子（RSS），抑制局部和全身RNA沉默。电泳迁移率转移实验表明，p24G1与双链21核苷酸小干扰RNA （siRNA）结合，而siRNA的结合是其RSS活性的必要条件，但不是充分条件。p24G1定位于细胞核内，可通过其氨基酸10 - 210区进行自相互作用。p24G1在移动到细胞核之前与输入蛋白α1相互作用需要二聚化，但其siRNA结合和RSS活性不需要二聚化。p24G1在双pGD载体或基于马铃薯病毒x的载体上的表达在烟草物种中引起了强烈的过敏反应，表明p24G1可能是发病的一个因素。此外，p24G1在发病机制中的功能需要其RSS活性、二聚化和核定位。此外，氨基酸122-139区域在p24G1的核输入、siRNA结合、沉默抑制和致病活性中起着至关重要的作用。这些结果有助于我们理解glrav1感染的分子机制。
Grapevine leafroll-associated virus 1 (GLRaV-1) is a major pathogen associated with grapevine leafroll disease. However, the molecular mechanisms underlying GLRaV-1 interactions with plant cells are unclear. Using Agrobacterium infiltration-mediated RNA-silencing assays, we demonstrated that GLRaV-1 p24 protein (p24G1) acts as an RNA-silencing suppressor (RSS), inhibiting local and systemic RNA silencing. Electrophoretic mobility shift assays showed that p24G1 binds double-stranded 21-nucleotide small interfering RNA (siRNA), and that siRNA binding is required but not sufficient for its RSS activity. p24G1 localizes in the nucleus and can self-interact through its amino acid 10 to 210 region. Dimerization is needed for p24G1 interaction with importin α1 before moving to the nucleus, but is not required for its siRNA binding and RSS activity. Expression of p24G1 from a binary pGD vector or potato virus X-based vector elicited a strong hypersensitive response in Nicotiana species, indicating that p24G1 may be a factor in pathogenesis. Furthermore, p24G1 function in pathogenesis required its RSS activity, dimerization and nuclear localization. In addition, the region of amino acids 122–139 played a crucial role in the nuclear import, siRNA binding, silencing suppression and pathogenic activity of p24G1. These results contribute to our understanding of the molecular mechanisms underlying GLRaV-1 infection.
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