Understanding DAWDLE Function in miRNA and siRNA Biogenesis

了解 DAWDLE 在 miRNA 和 siRNA 生物发生中的功能

• 批准号: 1121193
• 负责人: Bin Yu
• 金额: $ 49.95万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1121193&HistoricalAwards=false
• 关键词: Understanding; DAWDLE; Function; miRNA; siRNA

Intellectual Merit: Small RNAs, including microRNAs (miRNAs) and small interfering RNAs (siRNAs), are key regulators of many biological processes. Several RNA-binding proteins involved in general RNA metabolism are known to play important roles in regulating miRNA biogenesis. A challenge is to understand how RNA binding proteins affect miRNA biogenesis, as well as how the miRNA processing machinery interconnects with general RNA metabolism. This research targets these knowledge gaps. Preliminary studies demonstrated that DAWDLE (DDL), an RNA binding protein from Arabidopsis, is required for the accumulation of a subset of miRNAs and siRNAs. DDL interacts with pri-miRNA (precursor of miRNAs) and DCL1 (the processor of pri-miRNA) and positively regulates pri-miRNA processing. We hypothesize that DDL facilitates DCL1 to access and/or process a subset of pri-miRNAs. Specific objectives are to: 1) Analyze the role of DDL in miRNA biogenesis; 2) Analyze the function of protein domains of DDL in pri-miRNA processing; and 3) Analyze the role of DDL in siRNA biogenesis. This research will advance knowledge of processes leading to small RNA accumulation, which in turn will improve future efforts at elucidating the biological processes involving small RNAs and to refine small RNA-based technology that has been used to improve agricultural production and human health. Broader Impacts: This research will provide training for a postdoctoral researcher, a graduate student, undergraduate students, and high school students. The PI is a faculty sponsor/mentor for students participating in the Nebraska EPSCoR-supported Young Nebraska Scientists program, which fosters research exposure and participation by underrepresented minority high school students, and in the University of Nebraska-Lincoln Undergraduate Creative Activities and Research Experiences (UCARE) program. In addition, the PI pursues active collaborations with faculty at local liberal arts colleges to help reach students that may not have ready opportunity for research experiences. The PI is also integrating his latest research into a new course, "RNA Silencing", offered to upper level undergraduate and graduate students.

小RNA，包括microRNA（miRNAs）和小干扰RNA（siRNAs），是许多生物过程的关键调节因子。已知参与一般RNA代谢的几种RNA结合蛋白在调节miRNA生物合成中起重要作用。一个挑战是了解RNA结合蛋白如何影响miRNA生物合成，以及miRNA加工机制如何与一般RNA代谢相互联系。这项研究针对这些知识差距。初步研究表明，来自拟南芥的RNA结合蛋白DAWDLE（DAWDLE）是一种miRNA和siRNA亚类积累所必需的蛋白。miRNA与pri-miRNA（miRNA的前体）和DCL 1（pri-miRNA的加工者）相互作用，并正向调节pri-miRNA的加工。我们假设，miR-1促进DCL 1访问和/或处理pri-miRNAs的子集。具体目标是：1）分析siRNA在miRNA生物合成中的作用; 2）分析siRNA的蛋白结构域在pri-miRNA加工中的功能; 3）分析siRNA在siRNA生物合成中的作用。这项研究将推进导致小RNA积累的过程的知识，这反过来又将改善未来阐明涉及小RNA的生物过程的努力，并改进已用于改善农业生产和人类健康的基于小RNA的技术。更广泛的影响：本研究将为博士后研究员、研究生、本科生和高中生提供培训。PI是一个教师赞助商/导师的学生参加内布拉斯加州EPSCoR支持的年轻内布拉斯加州科学家计划，促进研究曝光和参与人数不足的少数民族高中学生，并在内布拉斯加州林肯大学本科生创意活动和研究经验（UCARE）计划。此外，PI寻求与当地文理学院的教师积极合作，以帮助那些可能没有现成的研究经验机会的学生。PI还将他的最新研究整合到一门新课程“RNA沉默”中，提供给高水平的本科生和研究生。