REGULATION OF POST-TRANSCRIPTIONAL GENE SILENCING

转录后基因沉默的调控

• 批准号: 6266319
• 负责人: VICKI VANCE
• 金额: $ 20.95万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6266319
• 关键词: Arabidopsis; RNA virus; calmodulin; endopeptidases; gene induction /repression; gene mutation; genetic mapping; genetic regulation; microarray technology; plant genetics; plant proteins; posttranscriptional RNA processing; protein protein interaction; virus protein; yeast two hybrid system

DESCRIPTION (from the application): Post-transcriptional gene silencing (PTGS) is an ancient eukaryotic regulatory mechanism in which a particular RNA sequence is targeted and destroyed. Although PTGS occurs in diverse organisms, the cellular components of the pathway and its regulation are not well understood. The helper component proteinase (HG-Pro) of plant potyviruses has been shown to suppress PTGS in plants. Using HG-Pro as bait in a yeast two-hybrid system, we have identified a novel calmodulin-related protein (termed rgsCaM) that interacts with this viral suppressor of PTGS. rgs-CaM, like HG-Pro itself, suppresses both initiation and maintenance of PTGS. Our hypothesis is that HC-Pro-suppression of PTGS is mediated by its interaction with rgs-CaM. Here we propose to exploit HC-Pro and rgs-CaM as tools to dissect the mechanism of silencing in plants. The first four aims of the proposal focus on the role of rgs-CaM in PTGS, identifying domains of the protein required to suppress PTGS as well as those involved in the rgs-CaM/HC-Pro interaction. Several approaches will be used to interfere with expression of rgs-CaM and the effect of these manipulations on PTGS and its suppression by HC-Pro will be assayed. The third aim addresses mechanistic questions, determining how and where HC-Pro and rgs-CaM act with regard to known steps in PTGS. The last two aims focus on the role of other plant proteins in the HC-Pro-mediated suppression of PTGS. We will extend investigations of three other HC-Pro-interacting proteins using the approaches that successfully identified rgs-CaM as a regulator of PTGS. Gene array technology will be used to identify genes that are regulated in response to PTGS or its reversal by HC-Pro. Finally, we will exploit the model genetic organism Arabidopsis thalianato screen for mutants that interfere with suppression of PTGS in response to HC-Pro. The plant silencing system serves as a model to understand similar pathways in animals, and has potential to be exploited for gene therapy applications and manipulation of gene expression. Given the antiviral nature of gene silencing in plants, understanding PTGS in plants could well lead to the development of antiviral strategies in humans.

描述（来自应用程序）：转录后基因沉默（PTGS） 是一种古老的真核调节机制，其中特定的RNA 序列针对和破坏。尽管PTG发生在多种生物中，但 途径的细胞成分及其调节不好 理解。植物维病毒的辅助成分蛋白酶（HG-Pro）具有 已显示可抑制植物中的PTG。在酵母中使用HG-Pro作为诱饵 两个杂交系统，我们已经确定了一种新型的钙调蛋白相关蛋白 （称为RGSCAM）与PTG的这种病毒抑制剂相互作用。 RGS-CAM， 像HG-Pro本身一样，可以抑制PTG的启动和维护。我们的 假设是PTG的HC-Pro抑制是由其相互作用介导的 带有RGS-CAM。在这里，我们建议利用HC-Pro和RGS-CAM作为剖析工具 植物中沉默的机制。提案重点的前四个目标 关于RGS-CAM在PTG中的作用，识别所需的蛋白质域 抑制PTG以及参与RGS-CAM/HC-PRO相互作用的PTG。 几种方法将用于干扰RGS-CAM的表达和 这些操作对PTG的影响及其抑制HC-Pro将是 测定。第三个目的解决了机械问题，确定了如何和 HC-Pro和RGS-CAM在PTG中的已知步骤。最后两个 目的是关注其他植物蛋白在HC-Pro介导的作用 抑制PTG。我们将扩大对其他三个的调查 使用成功鉴定的方法的HC-Pro相互作用蛋白 RGS-CAM作为PTG的调节剂。基因阵列技术将用于识别 响应于PTG或其逆转而受到HC-Pro的逆转的基因。 最后，我们将利用模型遗传生物拟南芥拟南芥 筛选出干扰PTG响应PTG的突变体 HC-Pro。植物沉默系统是理解类似的模型 动物的途径，并且有可能被利用用于基因治疗 基因表达的应用和操纵。鉴于抗病毒的性质 植物中的基因沉默，了解植物中的PTG可能会导致 制定人类的抗病毒策略。