中文摘要：黄瓜花叶病毒（CMV）因其分布广泛、复制水平高、基因组结构简单等特点，成为植物与病毒蛋白交互作用和机制研究的模式病毒。2b蛋白是由CMV编码的多功能蛋白，可抑制基于RNA沉默的抗病毒防御，有助于CMV在寄主植物中的毒力表达。本研究通过酵母双杂交系统从拟南芥cDNA文库筛选出12种CMV LS2b相互作用寄主蛋白。从寄主蛋白中选择30S核糖体亚基蛋白S11 (RPS11)进一步研究。研究结果表明，病毒LS2b蛋白的功能会非常显著地受到其与寄主RPS11蛋白之间交互作用的影响。
外文摘要：Cucumber mosaic virus (CMV) is a model virus for plant–virus protein interaction and mechanism research because of its wide distribution, high-level of replication and simple genome structure. The 2b protein is a multifunctional protein encoded by CMV that suppresses RNA silencing-based antiviral defense and contributes to CMV virulence in host plants. In this report, 12 host proteins were identified as CMV LS2b binding partners using the yeast two-hybrid screen system from the Arabidopsis thaliana cDNA library. Among the host proteins, 30S ribosomal subunit protein S11 (RPS11) was selected for further studies. The interaction between LS2b and full-length RPS11 was confirmed using the yeast two-hybrid system. Bimolecular fluorescence complementation (BIFC) assays observed by confocal laser microscopy and Glutathione S-transferase (GST) pull-down assays were used to verify the interaction between endogenous NbRPS11 and viral CMVLS2b both in vivo and in vitro. TRV-based gene silencing vector was used to knockdown NbRPS11 transcription, and immunoblot analysis revealed a decline in infectious viral RNA replication and a decrease in CMV infection in RPS11 down-regulated Nicotiana benthamiana plants. Thus, the knockdown of RPS11 likely inhibited CMV replication and accumulation. The gene silencing suppressor activity of CMV2b protein was reduced by the RPS11 knockdown. This study demonstrated that the function of viral LS2b protein was remarkably affected by the interaction with host RPS11 protein.
外文关键词：2B PROTEIN; PLANT; RESISTANCE; TOBACCO; ACCUMULATION; MOVEMENT; VECTOR
作者：R Wang ， Z Du ， Z Bai ， Z Liang
作者单位：西北农林科技大学
期刊名称：Plos One
期刊影响因子：2.8059
出版年份：2017
出版刊次：8
点击下载：内源性30S核糖体亚基蛋白S11和黄瓜花叶病毒LS2b蛋白之间的交互作用对病毒复制、侵染以及基因沉默抑制子活性的影响研究