Structural Biophysics of HIV Splicing and Transcriptional Control

HIV剪接和转录控制的结构生物物理学

• 批准号: 10399137
• 负责人: Blanton Smith Tolbert
• 金额: $ 15.94万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 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/10399137
• 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

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：尽管抗逆转录病毒疗法进展，艾滋病毒仍对人类构成严重威胁 卫生和全球经济。了解控制HIV基因的基本机制方面的知识差距 表达呈现出确定治疗干预或策略的新目标的关键障碍 teract病毒潜伏期。在此提案中，我们努力弥合有关RNA构成贡献的那些差距 呼吸动力学对艾滋病毒的剪接和转录控制。我们认为，艾滋病毒使用结构化的RNA元素 指导特定RNA蛋白（RNP）复合物的合作组装，从而调节剪接。这 RNA介导的合作性的类型是一种创新的概念，它解释了为什么HIV保存RNA结构 在其剪接站点附近。这些结构及其构象动态可以积极或负面 合作蛋白 - 蛋白质相互作用；我们试图了解这种机制的生物物理性质。我们 目前的有力证据表明，围绕HIV剪接位点A2和A3的RNA相互作用在本质上调节 VPR和TAT的水平。那些RNA结构内的突变抑制了通过deregu-抑制HIV复制 lating RNA剪接。我们进一步争辩说，类似的RNA介导的合作机制决定了前提 细胞P-TEFB从7SK snRNA释放并刺激HIV转录的帐篷 伸长。为了测试RNA介导的合作性影响HIV核基因表达的程度，我们 已经制定了一项技术创新的研究计划，该计划将尖端生物物理和生物化学融为一体 提取有关宿主病毒RNP复合物的机理原理的ICAL技术。我们将实施此专业 克通过采用两个广泛概念的特定目的：1。病毒RNA和RNP的分子相互作用 有助于HIV剪接和2。7SK SNRNA及其不同RNP的分子相互作用 HIV转录。该提案的成功有望将目光前所未有的见面交付到结构上 控制HIV基因表达的RNA-RNA和蛋白RNA相互作用的动力学，并且可以告知 细节介绍了有助于潜伏期的分子机制。反过来，这将为识别 治愈策略的新目标。