Functional role of RNA structure and m6A modification in viral genomes

RNA结构和m6A修饰在病毒基因组中的功能作用

基本信息

批准号：
10458770
负责人：
Mark A Boerneke
金额：
$ 12.1万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10458770
关键词：
Affinity Binding Binding Proteins Bioinformatics Biological Biological Assay Biological Models Biology Cell Culture Techniques Cells Chemicals Child Complex Consensus Dengue Dengue Virus Disease Educational workshop Elements Enterovirus Enterovirus 68 Environment Epigenetic Process Family Foundations Gene Expression Regulation Genetic Transcription Genome Goals Health Higher Order Chromatin Structure Human Immunologic Factors Infection Instruction Laboratories Life Cycle Stages Link Maps Mass Spectrum Analysis Mediating Mentors Metabolism Modeling Modification Mutation Nucleotides Phase Play Positioning Attribute Protein Biosynthesis Proteins RNA RNA Conformation RNA Viruses RNA-Protein Interaction Regulation Research Resolution Respiratory Disease Role Scientist Site Solid Structure Structure-Activity Relationship Techniques Therapeutic Training Vaccine Therapy Vaccines Viral Viral Genome Virion Virus Virus Diseases Virus Replication Work acute flaccid myelitis base career design disorder control genome-wide improved mortality mosquito-borne mutant nervous system disorder next generation novel prevent programs respiratory symposium vaccination strategy viral RNA virology virus culture

项目摘要

PROJECT SUMMARY/ ABSTRACT RNA viruses encode the information required to usurp cellular metabolism and gene regulation and to enable their own replication in two ways: in the linear sequence of their RNA genomes and in complex higher order structures. Although structured RNA elements are pervasive throughout viral genomes and have complex regulatory effects on all stages of the virus life cycle, little is known about the extent to which RNA structures occur across viral genomes or how critical structures function mechanistically. Post-transcriptional RNA genome chemical modifications such as N6-methyladenosine (m6A) are regulators of infection in diverse viruses and can have profound impacts on, or be impacted by, RNA structure. However, the link between RNA structure- and m6A-mediated regulation of viral infectivity remains uncharacterized because we lack a comprehensive structural understanding of RNA genomes and studies mapping m6A modifications have been imprecise. During the mentored phase of this proposal, I will gain new training in virology, RNA epigenetics, and RNA chemical biology to define how RNA elements and m6A modifications regulate viral replication and infection. In Aim 1, I will characterize RNA structure interrelationships with the m6A post-transcriptional RNA chemical modification by creating high-resolution RNA structure models and m6A modification site maps for the dengue virus (DENV) and the respiratory enterovirus EV-D68 RNA genomes. No broadly effective vaccines or therapeutics are available to prevent or treat the serious infections caused by these two single-stranded, positive-sense RNA viruses from distinct virus families. Through Aim 2, I will establish the functional importance of m6A-related RNA genome structures in DENV and EV-D68 viral life cycle stages using cell culture-based virus functional assays. Finally, in Aim 3, I will define structure-function relationships in m6A-related RNA structures in DENV and EV-D68 genomes by evaluating dynamic changes in RNA structure, m6A modification, and protein binding. Together, these studies will reveal the complex roles that interrelationships between RNA structure and m6A modification play in regulating the life cycles of diverse RNA viruses and will identify novel RNA regulatory motifs that might be exploited in the design of anti-DENV and anti-EV-D68 therapeutics and vaccination strategies. I have assembled an expert team of mentors and collaborators and plan to attend workshops, seminars, and conferences that will result in the training necessary to achieve the research goals proposed here and to transition into a successful career as an independent research scientist. The excellent training environments in the Weeks and Horner laboratories, along with hands-on training from my collaborator Dr. Cameron, will provide me with a solid foundation on which to build a successful independent research program.

项目摘要/摘要 RNA病毒编码篡夺细胞代谢和基因调节所需的信息，并启用他们自己的复制有两种方式：以其RNA基因组的线性序列和复杂的更高顺序结构。尽管结构化的RNA元素在整个病毒基因组中普遍存在，并且具有复杂对病毒生命周期的所有阶段的调节作用，关于RNA结构的程度知之甚少发生在病毒基因组或关键结构机械上的功能。转录后RNA基因组 N6-甲基拉丹氨酸（M6A）等化学修饰是各种病毒中感染的调节剂，并且可以对RNA结构产生深远影响或受到RNA结构的影响。但是，RNA结构与 M6A介导的病毒感染性调节仍然没有表征，因为我们缺乏全面的结构对RNA基因组和研究映射M6A修饰的理解已不精确。在该提案的指导阶段，我将获得病毒学，RNA表观遗传学和RNA化学生物学的新培训定义RNA元素和M6A修饰如何调节病毒复制和感染。在AIM 1中，我会通过M6A转录后RNA化学修饰来表征RNA结构的相互关系为登革热病毒（DENV）和呼吸道肠病毒EV-D68 RNA基因组。没有广泛有效的疫苗或治疗剂为了防止或治疗由这两种单链，正义的RNA病毒引起的严重感染不同的病毒家族。通过AIM 2，我将确定与M6A相关RNA基因组的功能重要性使用基于细胞培养的病毒功能分析的DENV和EV-D68病毒生命周期阶段中的结构。最后，在AIM 3中，我将在DENV和EV-D68中的M6A相关RNA结构中定义结构功能关系通过评估RNA结构，M6A修饰和蛋白质结合的动态变化来评估基因组。一起，这些研究将揭示RNA结构和M6A修饰之间相互关系的复杂作用在调节各种RNA病毒的生命周期中发挥作用，并将确定可能在抗-DENV和Anti-EV-D68治疗和疫苗接种策略的设计中被利用。我有组建了一个由导师和合作者组成的专家团队，并计划参加研讨会，研讨会和会议将导致实现此处提出的研究目标所必需的培训过渡到独立研究科学家成功的职业生涯。出色的培训环境 Weeks和Horner Laboratories以及我的合作者Cameron博士的动手培训将提供我以坚实的基础来建立成功的独立研究计划。