Viral modulation of epitranscriptomic mechanisms

表观转录组机制的病毒调节

• 批准号: 10317748
• 负责人: Benjamin A Garcia
• 金额: $ 0.48万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-19 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10317748
• 关键词: Address; Adenovirus Infections; Adenoviruses; Affinity; Affinity Chromatography; Amino Acids; Antibodies; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Models; Biology; Cell Nucleus; Cell physiology; Cells; Cellular biology; Chemicals; DNA Viruses; Data; Detection; Disease; Double Stranded DNA Virus; Genetic; Genetic Transcription; Goals; Growth; Host Defense; Human; Immune response; Immune system; Individual; Infection; Inhibitor of Differentiation Proteins; Innate Immune Response; Knowledge; Life; Maps; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Methods; Modification; Nuclear; Nuclear Export; Oligonucleotides; Organ; Pathway interactions; Physiologic pulse; Post-Transcriptional RNA Processing; Process; Production; Proteins; Proteomics; RNA; RNA Processing; RNA Sequences; RNA Splicing; RNA analysis; RNA metabolism; RNA-Binding Proteins; RNA-Protein Interaction; Reader; Regulation; Respiratory Tract Infections; Role; Stable Isotope Labeling; Study models; System; Technology; Tertiary Protein Structure; Time; Translating; Translations; Untranslated RNA; Viral; Viral Genes; Virus; Virus Diseases; base; combat; epitranscriptome; epitranscriptomics; gene product; human disease; human pathogen; improved; new technology; protein complex; protein expression; tool; transcriptome sequencing; viral RNA; virus host interaction

PROJECT SUMMARY The overall goal of this proposal is to develop high-throughput mass spectrometry technologies applied to the study of post-transcriptional RNA modifications (PTrMs) and associations with RNA binding proteins (RBPs). Efficient RNA processing and protein translation requires interactions with numerous RNA binding proteins, and can be regulated by chemical RNA modifications. PTrMs have been implicated in such diverse processes as RNA splicing, nuclear export, stability, and translation. The mechanisms by which PTrMs control RNA fate has opened up a new field dubbed “Epitranscriptomics”. Although there are antibody approaches to detect some modifications, there is a need for orthogonal approaches for unbiased identification and analysis of PTrMs. Since small DNA viruses that replicate in the nucleus have both to employ cellular machinery to transcribe and translate their gene products, and also develop ways to counteract host defenses, these viruses harness and manipulate cellular RNA processing pathways. Virus infections thus provide elegant biological models to decipher how RNA transcription and its chemical modifications can be regulated and exploited to direct the host cell machinery towards production of viral progeny. Based on preliminary data generated by our collaborative team, our objectives in this proposal are to employ Adenovirus as a model system to study PTrMs on non-coding and messenger viral RNAs and how they are exploited to counter host defenses and promote efficient viral RNA processing and progeny production. Adenoviruses are large non-eveloped viruses and include over 50 distinct strains which elicit a wide range of effects in humans, from respiratory infections to life-threatening organ problems in people with weakened immune systems. Adenoviruses hijack the host cell machinery to express viral genes, and this is achieved by overtaking RNA-mediated processes. Our preliminary data show how Adenovirus infection exploits the m6A modification on RNA to promote splicing and also alters RNA-protein interactions. Here we will develop improved mass spectrometry (MS) technologies to quantitatively and comprehensively detect RNA modifications and RNA-protein interactions over a detailed time-course of infection. Our MS technology will be paired with cellular and genetic assays to determine how the modifications are utilized by Adenovirus to promote growth and counter host defenses in ways that culminate in human disease. These high-throughput unbiased MS-based technologies and approaches will be broadly applicable to enable new biology in virus-host interactions and epitranscriptomics.

项目概要 该提案的总体目标是开发应用于 研究转录后 RNA 修饰 (PTrM) 以及与 RNA 结合蛋白 (RBP) 的关联。 有效的 RNA 加工和蛋白质翻译需要与大量 RNA 结合蛋白相互作用，并且 可以通过化学 RNA 修饰来调节，PTrM 涉及多种过程，例如 RNA 剪接、核输出、稳定性和翻译 PTrM 控制 RNA 命运的机制。 尽管有抗体方法可以检测某些内容，但开辟了一个被称为“表观转录组学”的新领域。 由于修改，需要采用正交方法来公正地识别和分析 PTrM。 在细胞核中复制的小型 DNA 病毒必须利用细胞机器进行转录和翻译 这些病毒利用它们的基因产物，并开发出对抗宿主防御的方法 因此，病毒感染的细胞RNA加工途径提供了优雅的生物学模型来破译RNA的加工过程。 转录及其化学修饰可以被调节和利用来指导宿主细胞机制 根据我们的合作团队生成的初步数据，我们的病毒后代的生产。 该提案的目标是采用腺病毒作为模型系统来研究非编码和非编码的PTrMs。 信使病毒 RNA 以及如何利用它们来对抗宿主防御并促进有效的病毒 RNA 腺病毒是大型非进化病毒，包括 50 多种不同的病毒。 对人类产生广泛​​影响的菌株，从呼吸道感染到危及生命的器官 免疫系统较弱的人会遇到腺病毒劫持宿主细胞机制来表达的问题。 病毒基因，这是通过超越 RNA 介导的过程来实现的，我们的初步数据显示了如何实现。 腺病毒感染利用 RNA 上的 m6A 修饰来促进剪接并改变 RNA 蛋白 在这里，我们将开发改进的质谱（MS）技术来定量和分析相互作用。 全面检测感染的详细时间过程中的 RNA 修饰和 RNA-蛋白质相互作用。 我们的 MS 技术将与细胞和遗传分析相结合，以确定如何利用修饰 腺病毒促进生长并对抗宿主防御，最终导致人类疾病。 高通量、公正的基于 MS 的技术和方法将广泛适用于实现新的 病毒与宿主相互作用和表观转录组学的生物学。