Structural insights into RNA promoters for RNA polymerase II-catalyzed RNA-templated transcription

RNA 聚合酶 II 催化 RNA 模板转录的 RNA 启动子的结构见解

• 批准号: 2145967
• 负责人: Ying Wang
• 金额: $ 52.28万
• 依托单位: Mississippi State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2145967&HistoricalAwards=false
• 关键词: Structural; insights; into; RNA; promoters

This project will shed light on how a DNA-directed RNA polymerase, normally involved in transcription of cellular DNA, is also able to recognize RNA templates. To understand how transcription polymerases can recognize RNA templates, this project aims to determine the molecular basis of a regulatory element within a well-known RNA template (potato spindle tuber viroid) that is critical for recruiting the transcription enzymes. To achieve this goal, this project will employ a combination of molecular, biochemical, and bioinformatic approaches. The knowledge gained from this project may facilitate the development of RNA-based technologies for basic research as well as applications in biomedicine and agriculture, thus benefiting society in the long term. This project will also serve as a vehicle for training undergraduate and graduate students. Particularly, undergraduate students from Delta State University, located in one of the least affluent regions in the US and lacking the necessary facility for research training, will participate in summer research annually to gain research experience and to be prepared for STEM-related careers.Supported by published and preliminary data, a testable hypothesis is that potato spindle tuber viroid recruits RNA-specific transcription factor(s), thereby enhancing the efficiency of RNA polymerase II (Pol II)-catalyzed RNA-templated transcription. The RNA promoter region is defined by the binding region of a recently identified RNA-specific transcription factor, TFIIIA-7ZF. This project will map and characterize the molecular basis/structure of RNA promoters within both sense and antisense genomes of potato spindle tuber viroid and its relatives, using a combination of selective 2’-hydroxyl acylation analyzed by primer extension (SHAPE) analysis, functional mutagenesis, Pol II-based in vitro transcription assay, RNA footprinting assays, etc. The outcomes from this project will help better understand the nature of RNA promoters and potentially exploit RNA promoters for usage in basic science and its applications in agriculture. While working on this project, undergraduate students from two universities in the state of Mississippi will receive experience in basic research in molecular biology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目将阐明DNA指导的RNA聚合酶（通常参与细胞DNA的转录）如何也能够识别RNA模板。为了了解转录聚合酶如何识别RNA模板，该项目旨在确定对募集转录酶至关重要的众所周知的RNA模板（马铃薯纺锤块茎）中调节元件的分子基础。为了实现这一目标，该项目将采用分子，生化和生化方法的结合。从该项目中获得的知识可能支持基于RNA的基础研究技术以及生物医学和同意的应用，从而使社会从长远来看受益。该项目还将作为培训本科生和研究生的工具。 Particularly, undergraduate students from Delta State University, located in one of the least affecting regions in the US and lacking the necessary facility for research training, will participate in summer research annually to gain research experience and to be prepared for STEM-related careers.Supported by published and preliminary data, a testable hypothesis is that potato spindle tuber viroid reports RNA-specific transcription factor(s), thereby enhancing the efficiency of RNA聚合酶II（POL II）催化的RNA-诱导转录。 RNA启动子区域由最近鉴定出的RNA特异性转录因子TFIIIA-7ZF的结合区域定义。该项目将使用选择性的2'-羟基酰基（通过引物扩展）分析（形状）分析，功能性诱变，从基于POL II的Intercription Introlays Outna Introkny Outna Introk Provecter等组合，将在有意义的块状块茎和其亲属的意义和纺锤体基因组中绘制和表征RNA启动子的分子基础/结构，从而使RNA脚底较高的脚步均可在此分析，RNA的尺度等等，等等。促进者和潜在利用RNA启动子来用于基础科学及其在协议中的应用。在从事该项目的过程中，来自密西西比州两所大学的本科生将获得分子生物学基础研究的经验。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的评估来评估的。

项目成果

A nuclear import pathway exploited by pathogenic noncoding RNAs.

• DOI: 10.1093/plcell/koac210
• 发表时间: 2022-09-27
• 期刊: The Plant cell

Emerging value of the viroid model in molecular biology and beyond.

• DOI: 10.1016/j.virusres.2022.198730
• 发表时间: 2022-05
• 期刊: Virus research
• 影响因子: 5
• 作者: Ma J;Mudiyanselage SDD;Wang Y
• 通讯作者: Wang Y