Functional Role of BYDV 3' RNA Translation Enhancer Element

智力优势：几乎所有的信使核糖核酸都有很好的证据证明长范围的RNA-RNA相互作用。大多数细胞内的mRNAs都有一个5‘端的帽和一个3’端的聚(A)尾，它们在mRNA的翻译中起着关键的作用。然而，涉及内部核糖体进入位点和3‘RNA元件的相互作用也开始在细胞RNA中被发现，特别是在整体翻译受到影响的情况下。对于那些缺少帽结构或Poly(A)尾巴或两者兼而有之的mRNA(通常是病毒)，会发生与经典翻译模型的重大偏差。其中一种病毒是大麦黄矮病毒(BYDV)，它是大麦中分布最广、经济上最重要的病毒之一。BYDV在3‘非翻译区含有一种不寻常的RNA结构，该结构与5’非翻译区的茎环相互作用。这种长距离的相互作用促进了病毒mRNA的高效翻译，但其机制尚不清楚。这个项目将探索BYDV RNA与eIF4F的相互作用，eIF4F是一种控制翻译启动的限速步骤的蛋白质。核糖体与黄矮病病毒RNA结合的特异性也将通过“脚印”分析来确定。综上所述，这些数据将有助于更好地理解病毒mRNA是如何在有效的病毒感染中击败宿主细胞信使RNA的。更广泛的影响：这些实验将为从高中到研究生院的学生提供生化和定量测量方面的极好培训。将继续与当地一所高中和曼哈顿社区学院(一家拉美裔服务机构)的教职员工进行互动，以帮助改善教职员工及其学生的研究。过去参与国际和平研究所研究的许多学生都来自代表人数较少的少数族裔群体，少数族裔学生也将被包括在这个项目中。这些学生将为科学和技术领域的多样化和训练有素的劳动力做出贡献。另一项活动是重新设计高级实验课程，以包括基于探究的研究，并为更多的学生提供研究体验。这项研究将通过使用植物病毒模型来研究基因表达调控的关键步骤，从而产生广泛的科学影响。植物病毒病影响世界各地的大量粮食作物，并可对经济条件和粮食供应产生严重影响。这项研究的信息不仅有可能用来控制病毒，还有可能开发出生产所需蛋白质的系统，这些蛋白质对营养有益或经济有用。

Intellectual Merit: Long range RNA-RNA interactions are well documented for virtually all mRNA. Most cellular mRNAs employ a 5' cap and a 3' poly (A) tail, both of which play key roles in translation of the mRNA. However interactions involving internal ribosome entry sites and 3' RNA elements are also beginning to be identified in cellular RNAs, especially under conditions where overall translation is compromised. A major deviation from the classical model of translation occurs for those mRNAs (often viral) that lack either a cap structure, poly (A) tail or both. One such virus is barley yellow dwarf virus (BYDV), one of the most widespread and economically important viruses of barley. BYDV contains an unusual RNA structure in the 3' untranslated region of the mRNA that interacts with a stem-loop in the 5' untranslated region. This long-range interaction promotes efficient translation of the viral mRNA, but the mechanism is not well understood. This project will explore the interaction of the BYDV RNA with eIF4F, a protein that controls the rate-limiting step for translation initiation. The specificity of ribosome binding to the BYDV RNA will also be determined by "toeprint" analysis. Taken together, these data will lead to a better understanding of how viral mRNA is able to out-compete host cell messenger RNA for effective viral infection. Broader Impacts: These experiments will provide excellent training for students from high school through graduate school in both biochemical and quantitative measurements. Interactions will continue with faculty from a local high school and from Borough of Manhattan Community College (a Hispanic serving institution) to help improve research for faculty and their students. Many of the students involved in the PI's research in the past have been from underrepresented minority groups, and minority students will also be included in this project. These students will contribute to a diverse and well-trained workforce in science and technology. An additional activity will be the redesign of the advanced lab curriculum to include inquiry-based research and to provide a research experience for more students. The research will have broad scientific impact through its use of a plant virus model for studying a key step in regulation of gene expression. Plant viral diseases affect a significant number of food crops world-wide and can have severe impact both on economic conditions and food supply. There is a potential to use information from this research not only to control viruses but also to develop systems to produce desired proteins which are nutritionally beneficial or economically useful.