Collaborative Research: RNA Processing in Trypanosome Mitochondria

合作研究：锥虫线粒体中的 RNA 加工

• 批准号: 2140152
• 负责人: Jorge Cruz-Reyes
• 金额: $ 82.82万
• 依托单位: Texas A&M AgriLife Research
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2140152&HistoricalAwards=false
• 关键词: Collaborative; Research; RNA; Processing; Trypanosome

This project will study a distinctive mechanism of gene expression in ancient single-celled parasites called trypanosomes. In most organisms, RNA is copied from DNA and directs protein synthesis without changes in the code. However, in trypanosomal mitochondria (the ‘powerhouse’ of cells), RNA is edited extensively by addition or removal of specific nucleotides. In their complex life cycle, trypanosomes alternate between two very different hosts: humans (and other animals) and insects, which are the vectors of tranmission. The dramatically different environments that the parasites face in humans and insects demand rapid and large-scale metabolic and physiological changes, including in RNA editing. The crucial question of how the RNA editing mechanism is precisely regulated during parasite development remains unanswered and is the focus of this research. The project will have broad educational impact by providing interdisciplinary training opportunities for postdoctoral, graduate, undergraduate, and high school students, with a focus on groups traditionally underrepresented in the STEM disciplines.This project focuses on Trypanosoma brucei mRNA editing regulation and the key role of the REH2C complex that controls editing fidelity in editosomes. The research can shed new light on a question that has remained a mystery for decades: how is mRNA editing differentially controlled across the T. brucei life-cycle? Objective 1 will test the “partial-editing control” (PEC) model of exploiting partial editing to differentially inhibit mRNA maturation in two developmental forms of T. brucei. A recently developed pipeline for RNA-seq/bioinformatic analysis of edited RNAs will be employed for this purpose. Objective 2 will address the source of substrate specificity in editing, specifically to define the role of a rare DEAH-box helicase cofactor and its individual zinc fingers in the mechanism. Objective 3 will examine dynamic interactions of editing components and remodeling by the REH2C complex by elucidating the general organization of editosomes using genetic, biochemical (proximity labeling), and chemical (crosslinking-mass spectrometry) approaches. Overall, these studies will reveal new aspects of editing regulation in two life-cycle stages of the parasite, including editing fidelity, substrate specificity determinants, and editosome interactions.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.

该项目将研究称为锥虫的古老单细胞寄生虫中基因表达的独特机制。在大多数生物体中，RNA是从DNA复制而来的，并指导蛋白质合成而不改变密码。然而，在锥虫线粒体（细胞的“发电站”）中，RNA通过添加或去除特定核苷酸进行广泛编辑。在其复杂的生命周期中，锥虫在两个非常不同的宿主之间交替：人类（和其他动物）和昆虫，这是传播的载体。寄生虫在人类和昆虫中所面临的截然不同的环境需要快速和大规模的代谢和生理变化，包括RNA编辑。在寄生虫发育过程中如何精确调节RNA编辑机制的关键问题仍然没有答案，这是本研究的重点。该项目将通过为博士后、研究生、本科生和高中生提供跨学科的培训机会，产生广泛的教育影响，重点关注传统上在STEM学科中代表性不足的群体。该项目重点关注布氏锥虫mRNA编辑调控和REH 2C复合体在编辑体中控制编辑保真度的关键作用。这项研究可以为一个几十年来一直是个谜的问题提供新的线索：mRNA编辑是如何在T细胞中受到差异控制的。布鲁氏菌的生命周期？目的1将在两种发育形式的T细胞中测试利用部分编辑差异抑制mRNA成熟的“部分编辑控制”（PEC）模型。布鲁塞。最近开发的用于编辑RNA的RNA-seq/生物信息学分析的管道将用于此目的。目标2将解决编辑中底物特异性的来源，特别是定义一种罕见的DEAH盒解旋酶辅因子及其单个锌指在机制中的作用。目标3将检查编辑组件和REH 2C复合物重塑的动态相互作用，通过使用遗传，生物化学（邻近标记）和化学（交联-质谱）方法阐明编辑体的一般组织。总的来说，这些研究将揭示新的方面的编辑调节在两个生命周期阶段的寄生虫，包括编辑保真度，底物特异性决定因素，和editosome interactions.This奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的影响审查标准的支持。