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Is the cucumber mosaic virus 2b silencing suppressor protein regulated by direct interactions with the viral 1a protein?

黄瓜花叶病毒 2b 沉默抑制蛋白是否通过与病毒 1a 蛋白的直接相互作用来调节？

基本信息

批准号：
2273242
负责人：
金额：
--
依托单位：
University of Cambridge
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2019
资助国家：
英国
起止时间：
2019 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=studentship-2273242
关键词：
cucumber mosaic virus 2b silencing

项目摘要

PhD project strategic theme: Bioscience for sustainable agriculture and foodProposed title for PhD: Using the cucumber mosaic virus 2b silencing suppressor protein and the viral 1a protein interaction to identify the relationship between silencing pathways and plant insect resistance mechanisms.The cucumber mosaic virus (CMV) 2b silencing suppressor protein inhibits RNA silencing mediated by short-interfering (si)RNAs (a potent antiviral mechanism) and micro(mi)RNA-regulated plant development. These two activities of the 2b protein are distinct: suppression of antiviral silencing depends on 2b's ability to bind double-stranded siRNA, while its effect on miRNA-regulated development depend largely on the interaction of 2b with Argonaut (AGO)1 protein. The effect of the 2b protein on an anti-insect defence system controlled by AGO1 are antagonized by another CMV protein the 1a protein, a replication-associated protein with no previously known connection to silencing. Recent evidence suggests that this effect is achieved through a direct interaction between the 2b and 1a viral proteins. The aim of this PhD is to characterise the interaction between 1a and 2b using 1a and 2b-derived wild-type and mutant constructs for protein binding studies including bimolecular fluorescence complementation and FLAG pull-downs. These techniques, along with fluorescent microscopy techniques, will also be used to identify the localisation and cell biology of the 1a and 2b viral proteins. This PhD will attempt to determine if 1a also affects the silencing suppressor activity of 2b or just its effects on miRNA pathways. Finally, this PhD will use 1a and 2b-derived wild-type and mutant constructs in aphid assays to assess the effects of disrupted silencing pathways on insect resistance in plants.

博士项目战略主题：生物科学促进可持续农业和食品利用黄瓜花叶病毒（CMV）2b沉默抑制蛋白（silencingsuppressorprotein，2b）和病毒1a蛋白的相互作用，研究沉默途径与植物抗虫机制的关系（一种有效的抗病毒机制）和微（mi）RNA调节的植物发育。2b蛋白的这两种活性是不同的：抗病毒沉默的抑制取决于2b结合双链siRNA的能力，而其对miRNA调节的发育的影响主要取决于2b与Argonaut（AGO）1蛋白的相互作用。2b蛋白对由AGO 1控制的抗昆虫防御系统的作用被另一种CMV蛋白1a蛋白拮抗，1a蛋白是一种复制相关蛋白，与先前已知的沉默无关。最近的证据表明，这种作用是通过2b和1a病毒蛋白之间的直接相互作用实现的。本博士学位的目的是使用1a和2b衍生的野生型和突变体构建体来研究1a和2b之间的相互作用，用于蛋白质结合研究，包括双分子荧光互补和FLAG下拉。这些技术，沿着荧光显微镜技术，也将被用来确定1a和2b病毒蛋白的定位和细胞生物学。这个博士将试图确定1a是否也影响2b的沉默抑制活性，或者只是它对miRNA通路的影响。最后，这个博士将使用1a和2b衍生的野生型和突变体构建体在蚜虫测定中评估破坏沉默途径对植物抗虫性的影响。