Analysis of a ribosome-binding 3' translational enhancer in a plus-strand RNA virus

知识价值。所有生物体，从病毒到人类，都必须通过蛋白质合成（翻译）过程从其基因组合成蛋白质。植物病毒特别有趣，因为它们必须在不杀死宿主细胞的情况下与宿主细胞竞争有限的翻译因子。对植物病毒的研究表明，RNA基因组最远端的翻译起始位置（3'端）是吸引翻译成分进入基因组的关键区域，从而大大增强了翻译。最近发现，一种带有RNA基因组的病毒豌豆花叶病毒（Pea enation mosaic virus）的3'区含有一个翻译增强子，可以吸引核糖体（核心蛋白质合成机器），并可能通过帮助核糖体在基因组翻译后循环发挥作用。此外，该翻译增强子还参与与病毒另一端（5'端）附近的另一个元件的长距离连接。该项目将研究这种翻译增强子如何与核糖体相互作用，以及核糖体是否可以通过同时与3‘和5’ RNA元件相互作用来“桥接”病毒基因组的末端。由于病毒基因组不能同时复制和翻译，本研究还将调查病毒如何在这些不相容的活动之间切换。最后，该研究将探索寄主植物基因组中含有类似的核糖体结合元件的可能性，这些元件可以增强寄主rna的翻译。研究3‘区如何参与翻译的新范式将促使其他研究者正确地检测他们的基因或病毒中的3’翻译增强子，这将是翻译领域的变革。通过了解这些翻译增强子的功能，在设计新方法以在植物中生产更高水平的产品以改善农业性状时，应该有可能包括它们。更广泛的影响。这项研究将由研究生和博士后研究人员进行，他们将学习最先进的技术和模型构建，以促进他们作为科学家的发展。本科研究人员也将参与该项目的各个方面，了解科学发现的激动人心之处。马里兰州乔治王子县杜瓦尔高中（98%为少数民族）的大学先修课程高中学生和教师每周都会继续对基于研究的发现充满热情，有几名学生已经被顶尖学院/大学录取，主修STEM科目。首席研究员还将继续有效地向公众代表科学和科学家，最近担任探索电视连续剧的顾问，并参加托莱多大学周六上午的科学节目等活动。这项研究的具体例子也将通过首席研究员教授的入门生物学课程来激发和增强马里兰大学理科专业第一年学生的教育体验。

Intellectual Merit. All organisms, from viruses to man, must synthesize proteins from their genomes by the process of protein synthesis (translation). Plant viruses are especially intriguing, as they must compete with their host cells for limited translation factors without killing the cell. Research on plant viruses has revealed that the end of the RNA genome most distal to where translation begins (the 3' end) is a critical region for attracting components of translation to the genome, leading to substantially enhanced translation. The 3' region of one virus with an RNA genome, Pea enation mosaic virus, was recently found to contain a translational enhancer that attracts ribosomes, the central protein synthesizing machines, and may function by helping ribosomes recycle following genome translation. In addition, this translational enhancer also participates in a long-distance joining with another element near the other end (the 5' end) of the virus. This project will examine how this translational enhancer interacts with ribosomes and whether ribosomes can "bridge" the ends of the viral genome by interacting simultaneously with both 3' and 5' RNA elements. Since viral genomes cannot be replicated and translated at the same time, this research will also investigate how the virus switches between these incompatible activities. Finally, the research will explore the possibility that the host plant's genome contains similar ribosome-binding elements that enhance translation of host RNAs. Developing new paradigms for how the 3' region participates in translation will prompt other investigators to correctly examine their genes or viruses for 3' translational enhancers, which will be transformative in the translation field. By understanding how these translational enhancers function, it should be possible to include them when designing new ways to produce higher levels of products in plants for improvement of agricultural traits. Broader Impacts. This research will be conducted by graduate and postdoctoral researchers, who will learn state-of-the art techniques and model-building to enhance their development as scientists. Undergraduate researchers will also participate in all aspects of this project, learning about the excitement of scientific discoveries. Enthusiasm for research-based discovery will continue to be conveyed weekly to Advanced Placement high school students and teachers at DuVal High School (98% minority) in Prince George's County, Maryland, with several students already benefiting by being accepted at top colleges/universities and majoring in STEM subjects. The principal investigator will also continue to effectively represent science and scientists to the public, most recently as a consultant for a Discovery TV Series and participation in activities such as the Saturday morning science program at the University of Toledo. Specific examples from this research will also be used to excite and enhance the education experience of students in their first year as science majors at the University of Maryland through an introductory biology course taught by the principal investigator.