权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Characterization of tRNA-derived RNA Fragments (tRFs) in Respiratory Syncytial Vi

呼吸合胞体 Vi 中 tRNA 衍生的 RNA 片段 (tRF) 的表征

基本信息

批准号：
8813852
负责人：
Xiaoyong Bao
金额：
$ 38.7万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8813852
关键词：
Address Antiviral Agents Attention Attenuated Attenuated Live Virus Vaccine Biochemistry and Cellular Biology Biogenesis Bioinformatics Biological Biological Markers Biological Process Biology Breathing Bronchiolitis Cells Cellular biology Child Collaborations Communities Complex Computer Analysis Cytoplasm Data Development Ectopic Expression Elements Employee Strikes Epithelial Cells Exhibits Failure Family Foundations Functional RNA Future Genes Genetic Transcription Genome Glutamic Acid-Specific tRNA Glycine decarboxylase Goals Health Human respiratory syncytial virus Immune response Immunoprecipitation In Vitro Infant Infection Information Centers Life Lower Respiratory Tract Infection Lung Maps Mediating Messenger RNA MicroRNAs Molecular Molecular Biology Molecular Profiling Molecular Virology Morbidity - disease rate Nucleotides Pattern Play Pneumonia Production Protein Biochemistry Proteins Public Health RNA RNA Biochemistry RNA-Induced Silencing Complex Regimen Regulation Research Resources Respiratory Syncytial Virus Infections Respiratory syncytial virus Respiratory syncytial virus RSV proteins Ribosomal RNA Role Small Interfering RNA Small Nucleolar RNA System Techniques Technology Testing The Sun Therapeutic Therapeutic Intervention Transfer RNA Vaccine Design Viral Virus Virus Diseases Virus Replication angiogenin apolipoprotein B mRNA editing enzyme apolipoprotein E receptor 2 base cellular targeting data integration design endonuclease experience human DICER1 protein innovation insight member mortality next generation sequencing novel novel therapeutics nuclease polypeptide positional cloning protein expression respiratory response structural biology therapeutic vaccine vaccine candidate virology virus host interaction

项目摘要

DESCRIPTION (provided by applicant): The discovery of small non-coding RNAs (sncRNAs) with regulatory functions is a recent breakthrough in biology. Among sncRNAs, microRNAs (miRNAs) and virus-derived sncRNAs have emerged as elements of critical importance in controlling viral replication and host responses to viral infection. However, the expression patterns and functional aspects of other types of sncRNAs after viral infection are completely unexplored. To define the expression patterns of sncRNAs, as well as to discover novel regulatory sncRNAs in response to viral infection, we applied next-generation sequencing (NGS) to cells infected with human respiratory syncytial virus (RSV), the most common respiratory mucosal virus that causes lung and airway infections in infants and young children, in comparison to uninfected cells. We found that RSV infection leads to abundant production of tRNA-derived RNA fragments (tRFs) that are approximately 30 nucleotides (nts) long and correspond to the 5'- half of mature tRNAs (tRF5). The induction of tRF5 is virus-specific and RSV replication-dependent. At least one member of these tRF5s, which is derived from tRNA-Glu-CTC (tRF5-GluCTC), decreases the level of target mRNAs in the cytoplasm. It also plays a critical role in RSV infection, as RSV replication is attenuated by its suppression, but promoted by its ectopic expression. The biogenesis of this tRF is also specific, and mediated by a particular endonuclease (angiogenin, ANG), and not by other nucleases. The central hypothesis of this project is that tRFs are not a random by-product of tRNA degradation, but rather are functional molecules. This central hypothesis will be addressed by identifying the targets of tRF5-GluCTC in RSV-infected airway epithelial cells (Aim 1), determining the viral factor(s) controlling the trans-silencing activity of tRFs (Aim 2), and defining 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. Of note, despite the public health importance of RSV, no effective therapeutic interventions or vaccines are available. Therefore, the overall goal of this project is to use a combination of molecular virology, protein and RNA biochemistry, cellular and structural biology techniques, to identify the molecular mechanisms underlying the regulatory effects of this novel tRF on RSV replication. The results of this project will provide an important new perspective and novel regulatory mechanisms to study the interactions between the host and RSV, and lay the foundation for the future development of biomarkers, therapeutic interventions, and vaccine designs based on the interaction of tRFs with their targets, both host and virus-derived, and the biogenic mechanisms of tRFs.

描述（由申请人提供）：具有调节功能的小非编码RNA（sncRNA）的发现是生物学的最新突破。在sncRNA中，微小RNA（miRNAs）和病毒衍生的sncRNA已经成为控制病毒复制和宿主对病毒感染的应答的至关重要的元件。然而，其他类型的sncRNA在病毒感染后的表达模式和功能方面是完全未知的。为了确定sncRNA的表达模式，以及发现新的调节sncRNA对病毒感染的反应，我们将下一代测序（NGS）应用于感染人呼吸道合胞病毒（RSV）的细胞，RSV是导致婴幼儿肺部和气道感染的最常见的呼吸道粘膜病毒，与未感染的细胞相比。我们发现RSV感染导致大量产生tRNA衍生的RNA片段（tRFs），其长度约为30个核苷酸（nt），对应于成熟tRNA（tRF 5）的5 '-一半。tRF 5的诱导具有病毒特异性和RSV复制依赖性。这些tRF 5中的至少一个成员，其来源于tRNA-Glu-CTC（tRF 5-GluCTC），降低细胞质中靶mRNA的水平。它还在RSV感染中起关键作用，因为RSV复制通过其抑制而减弱，但通过其异位表达而促进。这种tRF的生物发生也是特异性的，并且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。该项目的中心假设是tRFs不是tRNA降解的随机副产物，而是功能分子。将通过鉴定RSV感染的气道上皮细胞中tRF 5-GluCTC的靶点（目的1）、确定控制tRF反式沉默活性的病毒因子（目的2）和定义有助于ANG介导的tRF生物发生的病毒组分（目的3）来解决这一中心假设。初步数据表明，RSV蛋白N和P将分别成为目标2和3的重点。值得注意的是，尽管RSV对公共卫生具有重要意义，但目前尚无有效的治疗干预措施或疫苗。因此，本项目的总体目标是使用分子病毒学、蛋白质和RNA生物化学、细胞和结构生物学技术的组合，以确定这种新型tRF对RSV复制的调节作用的分子机制。该项目的结果将为研究宿主与RSV之间的相互作用提供重要的新视角和新的调控机制，并为未来基于tRFs与其靶点（宿主和病毒来源）的相互作用以及tRFs的生物学机制的生物标志物，治疗干预和疫苗设计的发展奠定基础。