Interplay of RNA Structural Motifs with Base Modifications

RNA 结构基序与碱基修饰的相互作用

基本信息

批准号：
10246857
负责人：
Cynthia J Burrows
金额：
$ 33.79万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10246857
关键词：
8-hydroxyguanosine Adenosine Alzheimer&apos s Disease Biology Cell physiology Chemicals Chemistry Consensus DNA Diabetes Mellitus Disease Flavivirus Foundations G-Quartets Genome Guanosine HIV Health Hemolysin Hepatitis B Human Inflammation Inflammatory Injury Kinetics Knowledge Location Malignant Neoplasms Measurement Messenger RNA Methods Methylation Mission Modification Molecular Monitor Nucleic Acid Regulatory Sequences Nucleic Acids Nucleosides Nutrient Oligonucleotides Oxidative Stress Play Proteins Pseudouridine Public Health RNA RNA Folding RNA-Directed DNA Polymerase Reading Reporting Research Research Project Grants Reverse Transcription Ribosomal RNA Role Science Signal Transduction Simian virus 40 Site Solvents Stress Structure Technology Temperature Testing Translations Traumatic Brain Injury United States National Institutes of Health Viral Viral Genome Virus Diseases Virus Replication Writing Zika Virus adduct age related base bisulfite innovation mechanical force methyl group methylation biomarker nanopore next generation sequencing oxidation single molecule stem tool transcriptome viral RNA

项目摘要

PROJECT SUMMARY Viral RNA, like the human transcriptome, is punctuated by infrequent but critical base modifications and non- Watson-Crick motifs. Many knowledge gaps exist in understanding where, when and why certain modifications such as pseudouridine (Ψ) and N6-methyl-adenosine (m6A) are enzymatically written onto mRNA. Similarly, guanosine-rich regions of viral RNA and the human transcriptome that may potentially fold to G-quadruplex motifs are conserved in regulatory regions controlling translation and viral replication for reasons that remain unclear. This research project hypothesizes that secondary structural motifs such as stem-loop structures and G-quadruplexes constitute the recognition sites for RNA modification. Additionally, these sites are hotspots for oxidative modification (8-oxo-7,8-dihydroguanosine, rOG) such as occurs during oxidative stress generated by viral infections. Thus, the project will examine the interplay of base modification (pseudouridinylation, guanosine oxidation and adenosine methylation) with secondary structural motifs in RNA. New innovative chemical biology tools will be developed to sequence long mRNA strands for folded structures by examining the ability of protein nanopores to thread and translocate folded or unfolded RNA. Similarly, base modifications will be identified using specific chemistries to amplify signals from base modification. The specific aims are to (1) investigate the sequence vs. structural motif of pseudouridine locations in ZIKV RNA, (2) sequence for rOG and correlate sites with secondary structure vs. solvent exposure, and (3) correlate G4 folds of ZIKV RNA with m6A. The human health relevance of this research is to provide foundational science for understanding the molecular choreography of mRNA, both human and viral, in order to advance health strategies combatting viral infection, cancer, and age-related disorders.

项目概要病毒 RNA 与人类转录组一样，不时出现不常见但关键的碱基修饰和非修饰。沃森-克里克主题在理解某些内容的地点、时间和原因方面存在许多知识差距。诸如假尿苷 (Ψ) 和 N6-甲基-腺苷 (m6A) 等修饰被酶促写入同样，病毒 RNA 和人类转录组中富含鸟苷的区域可能会折叠。 G-四链体基序在控制翻译和病毒复制的调控区域中是保守的该研究项目提出了二级结构主题，例如茎环结构和G四链体构成了RNA修饰的识别位点。这些位点是氧化修饰（8-oxo-7,8-二氢鸟苷，rOG）的热点，例如发生在因此，该项目将研究碱基修饰的相互作用。（假尿苷化、鸟苷氧化和腺苷甲基化）具有二级结构基序 RNA。将开发新的创新化学生物学工具来对折叠的长 mRNA 链进行测序。通过检查蛋白质纳米孔穿线和易位折叠或未折叠 RNA 的能力来研究结构。类似地，将使用特定的化学物质来识别碱基修饰，以放大来自碱基的信号。修饰的具体目的是（1）研究假尿苷的序列与结构基序。 ZIKV RNA 中的位置，(2) rOG 序列以及二级结构与溶剂的相关位点 (3) 将 ZIKV RNA 的 G4 折叠与 m6A 关联起来。这项研究的人类健康相关性是。为理解人类和人类 mRNA 的分子编排提供基础科学病毒，以推进对抗病毒感染、癌症和与年龄相关的疾病的健康策略。