RNA Uridylation in Trypanosomes

锥虫中的 RNA 尿苷化

基本信息

批准号：
10591650
负责人：
Ruslan Afasizhev
金额：
$ 64.18万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-05 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10591650
关键词：
Ablation Adaptor Signaling Protein Affect Africa South of the Sahara Antisense RNA Biogenesis Biology Catalytic Domain Cell Cycle Regulation Cells Code Complex Coupling Cryoelectron Microscopy Crystallization Crystallography DNA Degradation Pathway Development Disease Double-Stranded RNA Elements Enzymatic Biochemistry Enzymes Eukaryota Exhibits Exonuclease Gene Expression Genes Genetic Genetic Transcription Genetic Translation Goals Guide RNA Half-Life Hybrids Hydrolysis Insect Vectors Kinetoplast DNA Knowledge Laboratories Link Maternal Messenger RNA Mediating Messenger RNA Metabolic Methods MicroRNAs Mitochondria Mitochondrial RNA Modality Modeling Modification Molecular Molecular Machines Nuclear Nucleoproteins Parasites Parasitic infection Pathway interactions Perception Phosphodiesterase I Phylogenetic Analysis Polyadenylation Prevalence Process Protein Isoforms Proteins Public Health RNA RNA Cap-Binding Proteins RNA Decay RNA Degradation RNA Editing RNA Helicase RNA Processing RNA metabolism Reaction Resolution Ribonucleoproteins Roentgen Rays Role Route Shapes Site Specificity Structure Tail Testing Textiles Trans-Splicing Transcript Transcriptional Regulation Trypanosoma Trypanosoma brucei brucei Variant Work crosslink epitranscriptomics genetic information helicase hemoflagellate in vivo insertion/deletion mutation instrument mRNA Decay mRNA Transcript Degradation marginalized population mitochondrial messenger RNA novel operation particle pathogen programs protein complex spleen exonuclease structural biology synergism transcriptome uridylate

项目摘要

ABSTRACT Parasitic infections by Trypanosoma brucei spp. undermine public health and economy in Sub-Saharan Africa. These hemoflagellates are distinguished by the kinetoplast, a mitochondrial nucleoprotein body containing interlinked maxicircle and minicircle DNA. From a basic knowledge perspective, findings of polycistronic transcription, trans-splicing and RNA editing in trypanosomatids have profoundly influenced the conceptual fabric of RNA biology. Accordingly, ongoing work on U-insertion/deletion mitochondrial mRNA editing has produced pivotal breakthroughs, including discoveries of gRNA, the editosome, and the first RNA uridylating enzyme, KRET1 TUTase. Sustained efforts by the PI and other laboratories have expanded the uridylation’s breadth from a unique editing reaction into a compound transcriptome-shaping force prominent in most eukaryotes. Building on recently unraveled mitochondrial and cytosolic mRNA maturation and decay pathways, and progress in TUTase structural biology, we submit these Specific Aims: Aim 1 investigates the TUTase–3′-5′ exonuclease nexus underpinning mitochondrial RNA biogenesis and degradation. It focuses on mechanisms by which KRET1-containing molecular machines recognize diverse ribonucleoprotein particles, thus governing processing, functions, and metabolic fates of their cargo RNA. Molecular and Cryo-EM approaches will define transcript type-specific KRET1 complexes and determine their interactomes, in vivo RNA targets and near-atomic structures. We will analyze a novel RNA helicase, KREH3, as the potential link between massive antisense transcription of kinetoplast DNA and KRET1-mediated turnover. Aim 2 examines structural determinants and interactions enabling KRET2 and MEAT1 to recognize double- stranded RNA substrates, such as gRNA–mRNA hybrids. These programable mitochondrial TUTases execute U-insertion editing, which constitutes a Kinetoplastea-specific method of genetic information transfer. Aim 3 explores TbTUT3 and TbTUT4 as potential modulators of cytosolic mRNA decay, a principal conduit regulating nuclear gene expression in lieu of missing transcriptional control. By assessing transcriptome-wide U- tailed mRNAs prevalence and decay rates, we will test the coupling between mRNA 3′ uridylation and directional clearance. Beyond the parasite RNA metabolism, we are inquiring how the divergent auxiliary modalities confer orthogonal specificities and functions to a conserved and phylogenetically pervasive TUTase catalytic domain.

抽象的 Brucei spp锥虫感染。破坏撒哈拉以南非洲的公共卫生和经济。这些血红蛋白由动力盘（一种含有线粒体核蛋白体）区分相互链接的maxcircle和微圆DNA。从基本知识的角度来看锥形剂中的转录，变形和RNA编辑对概念结构有深远的影响 RNA生物学。彼此之间，正在进行的关于U-插入/缺失线粒体mRNA编辑的工作已产生关键突破，包括GRNA的发现，编辑体和第一个RNA尿苷酶， Kret1 tutase。 PI和其他实验室的持续努力已扩大了尿苷的广度在大多数真核生物中突出的复合转录组形状的独特编辑反应。建立在最近揭开的线粒体和胞质mRNA成熟和衰减途径的基础上，并进步在Tutase结构生物学中，我们提交了这些特定目的： AIM 1研究了tutase – 3'-5'外切核酸酶Nexus Nexus的线粒体RNA生物发生和降解。它着重于含KRET1的分子机的机制核糖核蛋白颗粒，因此是其货物RNA的加工，功能和代谢命运的处理。分子和冷冻EM方法将定义转录本类型特异性KRET1复合物并确定其相互作用，体内RNA靶标和近原子结构。我们将分析一种新型的RNA解旋酶Kreh3，作为动力学DNA的大量反义转录与KRET1介导的周转之间的潜在联系。 AIM 2考试结构决定者和相互作用，使Kret2和Meat1能够识别双重滞留的RNA底物，例如GRNA -MRNA杂种。这些可编程的线粒体教酶执行 U插入编辑，构成了遗传信息转移的动力塑料特异性方法。 AIM 3探索TBTUT3和TBTUT4作为主要导管的胞质mRNA衰变的潜在调节剂调节核基因表达代替缺失的转录控制。通过评估整个转录组的U- 尾mRNA患病率和衰减率，我们将测试mRNA 3'尿液和方向性之间的耦合清除。除了寄生虫RNA代谢之外，我们还在询问如何发散辅助模式会议正交的特异性和功能，用于配置和系统发育普遍的诱变酶催化结构域。