Structural Biophysics of HIV Splicing and Transcriptional Control

HIV剪接和转录控制的结构生物物理学

基本信息

批准号：
10034853
负责人：
Blanton Smith Tolbert
金额：
$ 35.25万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-19 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10034853
关键词：
Acquired Immunodeficiency Syndrome Adopted Binding Binding Sites Biochemical Biological Assay Biophysics Cells Chemicals Complex Data Elements Etiology Event Foundations Gene Expression Genetic Transcription HIV Health Heterogeneous-Nuclear Ribonucleoproteins Human Image Immunofluorescence Immunologic Knowledge Mediating Modeling Molecular Molecular Conformation Mutation Nature Nuclear Pathway interactions Phenotype Positive Transcriptional Elongation Factor B Process Property Proteins RNA RNA Binding RNA Splicing RNA-Binding Proteins RNA-Protein Interaction Regulatory Element Research Series Site Small Nuclear RNA Specificity Structural Biochemistry Structural Models Structural Protein Structure Sum Tars Techniques Testing Therapeutic Intervention Transcript Transcription Elongation Transcriptional Elongation Factors Transcriptional Regulation Transcriptional Silencer Elements Untranslated RNA Viral Viral Pathogenesis Virus Virus Latency Virus Replication Vision antiretroviral therapy base hnRNP protein A1 inhibitor/antagonist innovation insight new therapeutic target novel programs protein complex protein protein interaction reactivation from latency spatiotemporal success three dimensional structure viral RNA

项目摘要

Project abstract: Despite advances in antiretroviral therapy, HIV continues to pose serious threats to human health and global economies. Knowledge gaps in understanding the basic mechanisms that control HIV gene expression present critical barriers to identifying novel targets for therapeutic intervention or strategies to coun- teract viral latency. In this proposal, we endeavor to close those gaps concerning the contributions of RNA struc- tural dynamics to HIV splicing and transcriptional control. We posit that HIV uses structured RNA elements to direct the cooperative assembly of specific RNA-protein (RNP) complexes that in turn modulates splicing. This type of RNA-mediated cooperativity is an innovative concept that explains why HIV conserves RNA structures near its splice sites. Those structures and its conformational dynamics can either positively or negatively impart cooperative protein-protein interactions; we seek to understand the biophysical nature of such mechanisms. We present strong evidence that the RNA interactions surrounding HIV splice sites A2 and A3 intrinsically regulate the levels of vpr and tat, respectively. Mutations within those RNA structures inhibit HIV replication by deregu- lating RNA splicing. We further contend that similar RNA-mediated cooperative mechanisms determine the ex- tent to which the cellular P-TEFb is released from 7SK snRNA and made available to stimulate HIV transcription elongation. To test the extent to which RNA-mediated cooperativity influences HIV nuclear gene expression, we have developed a technically innovative research program that integrates cutting edge biophysical and biochem- ical techniques to extract mechanistic principles about host-virus RNP complexes. We will implement this pro- gram by adopting two broadly conceptual specific aims: 1. Molecular interactions of viral RNAs and RNPs that contribute to HIV splicing and 2. Molecular interactions of 7SK snRNA and its different RNPs that contribute to HIV transcription. The success of this proposal promises to deliver unprecedented in-sights into the structural dynamics of RNA-RNA and protein-RNA interactions that control HIV gene expression and it may inform on details into molecular mechanisms that contribute to latency. This will in turn provide a foundation for identifying novel targets for cure strategies.

项目摘要：尽管抗逆转录病毒治疗取得了进展，但艾滋病毒仍然对人类构成严重威胁健康和全球经济。了解控制 HIV 基因的基本机制方面的知识差距表达对确定治疗干预的新靶点或应对策略构成了关键障碍病毒潜伏期。在本提案中，我们努力缩小有关 RNA 结构贡献的差距。 HIV剪接和转录控制的自然动力学。我们假设 HIV 使用结构化 RNA 元件来指导特定 RNA-蛋白质 (RNP) 复合物的协同组装，进而调节剪接。这 RNA 介导的协同性类型是一个创新概念，解释了 HIV 为何保留 RNA 结构靠近其剪接位点。这些结构及其构象动力学可以产生积极或消极的影响蛋白质-蛋白质之间的协同相互作用；我们寻求了解此类机制的生物物理性质。我们强有力的证据表明 HIV 剪接位点 A2 和 A3 周围的 RNA 相互作用本质上调节分别是 vpr 和 tat 的水平。这些 RNA 结构内的突变通过 deregu 抑制 HIV 复制延迟RNA剪接。我们进一步认为，类似的 RNA 介导的合作机制决定了前 7SK snRNA 释放细胞 P-TEFb 并可刺激 HIV 转录伸长。为了测试 RNA 介导的协同性影响 HIV 核基因表达的程度，我们开发了一项技术创新的研究计划，整合了尖端的生物物理和生物化学- 提取宿主-病毒 RNP 复合物机制原理的技术。我们将实施这一亲 gram 通过采用两个广泛的概念性具体目标： 1. 病毒 RNA 和 RNP 的分子相互作用 2. 7SK snRNA 及其不同 RNP 的分子相互作用，有助于 HIV 剪接 HIV转录。该提案的成功有望为结构性问题提供前所未有的见解。控制 HIV 基因表达的 RNA-RNA 和蛋白质-RNA 相互作用的动力学，它可能提供信息详细了解导致延迟的分子机制。这将为识别提供基础治疗策略的新目标。