tRNA-derived RNA Fragments (tRFs) and their Functions in Respiratory Syncytial Virus (RSV) Infection

tRNA 衍生的 RNA 片段 (tRF) 及其在呼吸道合胞病毒 (RSV) 感染中的功能

• 批准号: 9384979
• 负责人: Xiaoyong Bao
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9384979
• 关键词: Algorithms; Antiviral Agents; Biochemistry and Cellular Biology; Biogenesis; Bioinformatics; Biological; Biological Process; Bronchiolitis; Cellular biology; Cleaved cell; Code; Collaborations; Communicable Diseases; Communities; Complex; Computer Analysis; Data; Development; Epithelial Cells; Exhibits; Expression Profiling; Failure; Family; Foundations; Future; Gene Expression Regulation; Genes; Genetic Transcription; Glycine decarboxylase; Goals; Human; Human Genetics; Immune response; Immunoprecipitation; Infant; Knowledge; Length; Lower Respiratory Tract Infection; Lower respiratory tract structure; Lung; Malignant Neoplasms; Mediating; Methodology; MicroRNAs; Molecular; Molecular Biology; Molecular Virology; Morbidity - disease rate; Nucleotides; Organism; Play; Pneumonia; Preventive; Prokaryotic Cells; Protein Biochemistry; Proteins; Public Health; Publications; Publishing; RNA; RNA Biochemistry; RNA-Induced Silencing Complex; Regulation; Regulator Genes; Research; Resources; Respiratory Syncytial Virus Infections; Respiratory syncytial virus; Respiratory syncytial virus RSV proteins; Role; System; Techniques; Testing; The Sun; Therapeutic; Transfer RNA; Untranslated RNA; Vaccines; Viral; Virus Replication; angiogenin; apolipoprotein B mRNA editing enzyme; apolipoprotein E receptor 2; biological systems; cellular targeting; data integration; design; early childhood; endonuclease; experience; experimental study; human disease; in vitro Assay; infancy; innovation; insight; mortality; nervous system disorder; novel; novel strategies; nuclease; polypeptide; protein expression; public health relevance; response; reverse genetics; structural biology; virology; virus host interaction

 DESCRIPTION (provided by applicant): Recent discoveries on small non-coding RNAs (sncRNAs) have significantly advanced human genetics and molecular biology, largely due to the identification of a fundamental role of sncRNAs as gene regulators. tRNA- derived RNA Fragments (tRFs) represent a recently discovered sncRNA family that is quickly being recognized as ubiquitously expressed in organisms ranging from prokaryotes to humans; however, tRFs' biological functions and the mechanism(s) underlying them are largely unknown. Respiratory syncytial virus (RSV) is the single most common cause of lower respiratory tract illness in infants. Our recent publication has shown that the sncRNAs most highly induced by RSV belong to the tRF family. Notably, at least the two most abundant RSV-inducible tRFs have a gene trans-silencing function that is mechanistically distinct from that of miRNAs, and one of these two, called tRF5-GluCTC, promotes RSV replication and regulates genes at a post-transcriptional level. The biogenesis of this tRF is also specific, and mediated by a particular endonuclease (angiogenin, ANG), and not by other nucleases. However, the molecular mechanisms underlying the biological functions and induction of tRF5-GluCTC are not known. Our central hypothesis in this project is that the RSV-induced tRF5-GluCTC is not a random by-product of tRNA degradation, but rather a functional molecule important for host-RSV interactions. This project will focus on exploring the molecular mechanisms underlying the regulatory effects of tRF5-GluCTC on RSV replication, and the host responses thereto. Our experiments will identify the targets of tRF5-GluCTC in RSV- infected airway epithelial cells (Aim 1), determine the viral factor(s) controlling the trans-silencing activity of tRF5- GluCTC (Aim 2), and define the viral component(s) contributing to ANG-mediated biogenesis of tRFs (Aim 3). Preliminary data suggest that the RSV proteins N and P will be a focus of Aims 2 and 3, respectively. The overall goal of this project is to use a combination of molecular virology, protein and RNA biochemistry, and cellular and structural biology techniques to elucidate tRF-mediated regulatory mechanisms for both RSV replication and the host responses it elicits. This novel regulation should provide a key new perspective on RSV-host interactions and undoubtedly facilitate the discovery of new gene regulatory mechanisms in response to RSV infection. Our results on antiviral target identification, targeting mechanisms, and tRF biogenesis will also provide new insights important for the design of preventive and therapeutic strategies for RSV infection. Given the early stage of studies on tRFs, the knowledge and techniques obtained in this study on tRF5-GluCTC will be important for exploring the functions of other tRFs in various biological systems, and therefore will broadly benefit the research community.

 描述（由申请人提供）：最近关于小非编码 RNA (sncRNA) 的发现极大地促进了人类遗传学和分子生物学的发展，这主要是由于发现了 sncRNA 作为 tRNA 衍生的 RNA 片段 (tRF) 的基本作用。代表最近发现的 sncRNA 家族，该家族很快被认为在从原核生物到人类的生物体中普遍表达，然而，tRF 的生物学功能和呼吸道合胞病毒 (RSV) 是婴儿下呼吸道疾病的最常见原因，我们最近的出版物表明，由 RSV 诱导的 sncRNA 很大程度上属于 tRF 家族。 ，至少两个最丰富的 RSV 诱导型 tRF 具有基因反式沉默功能，其机制在机制上不同于 miRNA，而这两个中的一个称为tRF5-GluCTC 促进 RSV 复制并在转录后水平调节基因。该 tRF 的生物发生也是特异性的，由特定的核酸内切酶（血管生成素，ANG）介导，而不是由其他核酸酶介导。 tRF5-GluCTC 的生物学功能和诱导尚不清楚。我们在该项目中的中心假设是 RSV 诱导的 tRF5-GluCTC 未知。 tRF5-GluCTC 是 tRNA 降解的随机副产物，而是对宿主与 RSV 相互作用重要的功能分子。该项目将重点探索 tRF5-GluCTC 对 RSV 复制的调节作用的分子机制，以及宿主对此的反应。将鉴定 tRF5-GluCTC 在 RSV 感染的气道上皮细胞中的靶标（目标 1），确定控制 tRF5- 反式沉默活性的病毒因子GluCTC（目标 2），并定义有助于 ANG 介导的 tRF 生物发生的病毒成分（目标 3）。初步数据表明，RSV 蛋白 N 和 P 将分别是目标 2 和 3 的重点。该项目的目标是结合分子病毒学、蛋白质和 RNA 生物化学以及细胞和结构生物学技术来阐明 tRF 介导的 RSV 复制和这种新的调控应该为RSV-宿主相互作用提供一个关键的新视角，并且无疑有助于发现针对RSV感染的新基因调控机制。 还将为 RSV 感染的预防和治疗策略的设计提供重要的新见解。鉴于 tRF 研究处于早期阶段，本次 tRF5-GluCTC 研究中获得的知识和技术对于探索其他 tRF 的功能非常重要。各种生物系统，因此将广泛惠及研究界。