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鉴定和表征一种富含甘氨酸的 RNA 结合蛋白,它是来自Nicotiana glutinosa 的内源性 RNA 沉默抑制剂。

Identification and characterisation of a glycine-rich RNA-binding protein as an endogenous suppressor of RNA silencing from Nicotiana glutinosa.

机构信息

State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Tai'an, Shandong, China.

Institute of Immunology, Taishan Medical University, Tai'an, Shandong, China.

出版信息

Planta. 2019 Jun;249(6):1811-1822. doi: 10.1007/s00425-019-03122-5. Epub 2019 Mar 6.

Abstract

This study shows that NgRBP suppresses both local and systemic RNA silencing induced by sense- or double-stranded RNA, and the RNA binding activity is essential for its function. To counteract host defence, many plant viruses encode viral suppressors of RNA silencing targeting various stages of RNA silencing. There is increasing evidence that the plants also encode endogenous suppressors of RNA silencing (ESR) to regulate this pathway. In this study, using Agrobacterium infiltration assays, we characterized NgRBP, a glycine-rich RNA-binding protein from Nicotiana glutinosa, as an ESR. Our results indicated that NgRBP suppressed both local and systemic RNA silencing induced by sense- or double-stranded RNA. We also demonstrated that NgRBP could promote Potato Virus X (PVX) infection in N. benthamiana. NgRBP knockdown by virus-induced gene silencing enhanced PVX and Cucumber mosaic virus resistance in N. glutinosa. RNA immunoprecipitation and electrophoretic mobility shift assays showed that NgRBP bound to GFP mRNA, dsRNA rather than siRNA. These findings provide the evidence that NgRBP acts as an ESR and the RNA affinity of NgRBP plays the key role in its ESR activity. NgRBP responds to multiple signals such as ABA, MeJA, SA, and Tobacco mosaic virus infection. Therefore, it could participate in the regulation of gene expression under specific conditions.

摘要

本研究表明,NgRBP 可抑制正义链或双链 RNA 诱导的局部和系统 RNA 沉默,其 RNA 结合活性对其功能至关重要。为了对抗宿主防御,许多植物病毒编码靶向 RNA 沉默各个阶段的病毒 RNA 沉默抑制剂,越来越多的证据表明,植物还编码内源性 RNA 沉默抑制剂 (ESR) 来调节该途径。在本研究中,我们通过农杆菌浸润分析,将来自 Nicotiana glutinosa 的富含甘氨酸的 RNA 结合蛋白 NgRBP 鉴定为一种 ESR。我们的结果表明,NgRBP 可抑制正义链或双链 RNA 诱导的局部和系统 RNA 沉默。我们还证明 NgRBP 可以促进 Potato Virus X (PVX) 在 N. benthamiana 中的感染。通过病毒诱导的基因沉默使 NgRBP 敲低,增强了 N. glutinosa 对 PVX 和黄瓜花叶病毒的抗性。RNA 免疫沉淀和电泳迁移率变动分析表明,NgRBP 结合 GFP mRNA、dsRNA 而不是 siRNA。这些发现提供了证据表明 NgRBP 作为 ESR 发挥作用,并且 NgRBP 的 RNA 亲和力在其 ESR 活性中起关键作用。NgRBP 对 ABA、MeJA、SA 和烟草花叶病毒感染等多种信号做出响应。因此,它可以在特定条件下参与基因表达的调控。

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