Regulation of Non-Canonical Telomerase RNA

本研究将探索一种新的基于rna的机制，使植物能够对影响DNA的胁迫做出反应，并传递保护染色体完整性的信号，这对植物的生育、生长和疾病预防至关重要。该项目将为培育和种植更健康的植物提供宝贵的基础知识。该项目将把基础植物科学研究与下一代植物科学家的教学和培训结合起来。作为这一努力的一部分，将为博士后研究员和新教员举办实验室管理研讨会，并将开发有关这一主题的研究生课程。本项目的总体目标是阐明TER2的分子机制和调控，TER2是一种新的端粒酶相关的长链非编码RNA (lncRNA)，来自模式植物拟南芥。TER2是端粒酶的负调节因子，在DNA双链断裂的反应中迅速稳定。TER2的诱导抑制端粒酶的活性，并被假设促进DNA损伤的忠实修复。TER2也是生殖健康所必需的，其机制尚不清楚。生物化学、遗传、细胞生物学和计算策略的结合将用于研究TER2。目的1将定义调节TER2加工/代谢的关键结构元件，以识别TER2内的DNA损伤传感器和控制TER2稳定性的元件。目的2将通过检测TER2是否是Dicer 2切割的底物来研究小RNA加工机制在促进TER2周转中的作用。目的3将通过检验TER2突变体的生育相关缺陷是否反映了异常的雄性配子体发育，还是父亲表达的印迹基因的影响，来研究TER2在拟南芥繁殖中的作用。总之，这项工作的结果将为lncRNA调控建立新的范例，并增加对促进基因组稳定性的基于rna的途径的理解。

This study will investigate a novel RNA-based mechanism that allows plants to respond to stress that affects the DNA, and to transmit a signal that protects chromosome integrity, which is important for plant fertility, growth and disease prevention. This project will develop fundamental knowledge valuable for breeding and growing healthier plants. This project will integrate basic plant science research with the teaching and training of the next-generation of plant scientists. As part of this effort, laboratory management workshops for postdoctoral fellows and new faculty will be conducted, and a graduate course on this topic will be developed.The overarching goal of this project is to elucidate the molecular mechanism and regulation of TER2, a novel telomerase-associated long noncoding RNA (lncRNA) from the model plant Arabidopsis thaliana. TER2 is a negative regulator of telomerase that is rapidly stabilized in response to DNA double-strand breaks. Induction of TER2 represses telomerase activity and is hypothesized to promote faithful repair of DNA damage. TER2 is also necessary for reproductive fitness through an unknown mechanism. A combination of biochemical, genetic, cell biological and computational strategies will be used to study TER2. Objective 1 will define key structural elements that regulate TER2 processing/metabolism for the purpose of identifying the DNA damage sensor within TER2 and the elements that control TER2 stability. Objective 2 will investigate the role of small RNA processing machinery in promoting TER2 turnover by testing if TER2 is a substrate for cleavage by Dicer 2. Objective 3 will examine the contribution of TER2 in Arabidopsis reproduction by testing whether the fertility-related defects in ter2 mutants reflect aberrant male gametogenesis or the effect of a paternally expressed imprinted gene. Altogether, the results of this work will establish new paradigms for lncRNA regulation and increase understanding of RNA-based pathways that promote genome stability.