Structural Biophysics of HIV Alternative Splicing

HIV选择性剪接的结构生物物理学

基本信息

批准号：
8704955
负责人：
Blanton Smith Tolbert
金额：
$ 29.18万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8704955
关键词：
3&apos Splice Site Acquired Immunodeficiency Syndrome Adopted Affect Affinity Alternative Splicing Binding Binding Sites Biochemical Biological Assay Biophysics Calorimetry Complex DNA Sequence Rearrangement Elements Ensure Environment Event Exons Genome Goals HIV HIV Genome Health Higher Order Chromatin Structure Homeostasis In Vitro Integration Host Factors Intervention Knowledge Lead Light Maps Messenger RNA Methods Modeling Molecular Monitor N-terminal NMR Spectroscopy Pathway interactions Pattern Property RNA RNA Sequences RNA Splicing Recruitment Activity Regulation Regulatory Element Repression Research Resolution Site Site-Directed Mutagenesis Structure Therapeutic Intervention Thermodynamics Titrations Transcript Viral Virus Work acrosome stabilizing factor base biochemical model hnRNP A1 in vivo innovation insight new therapeutic target novel novel strategies public health relevance research study stem three dimensional structure viral RNA

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to develop an innovative structural, biophysical, and functional biochemical framework to advance our understanding of HIV splicing mechanisms. The HIV genome is synthesized as a single polycistronic RNA transcript that undergoes regulated splicing to ensure mRNA homeostasis. Aberrations in the viral splicing machinery impair HIV replication fidelity and in some cases render the virus less infectious. Thus, HIV splicing pathways represent novel targets for therapeutic intervention. Splicing decisions are determined by the dynamic assembly/disassembly of trans host factors with cis viral RNA control elements; however, the underlying molecular events that determine whether a splice site will be activated or repressed remain elusive. Knowledge of the physical and functional determinants of how one highly conserved splice site gets differentially regulated by two trans host factors offers the opportunit to define a mechanism. Hence, the immediate goal of this proposal is to identify the molecular principles that HIV uses to co-opt the mutually antagonistic factors, hnRNP A1 (A1) and ASF, in order to regulate its 3' splice site A7 (ssA7). Furthermore, A1 and ASF regulate other conserved HIV splice sites via poorly characterized mechanisms; thus, the work proposed here is likely to have broad implications on our understanding of HIV splicing in general and may lead to new approaches to thwart HIV replication. The specific aims of this project are to: (1) Determine the contribution of structure to target site selection by A1 and ASF, (2) Elucidate the bio-thermodynamic underpinnings of mutual antagonism between A1 and ASF, and (3) Investigate the molecular origins of cooperativity in ssA7 regulation.

描述（由申请人提供）：该提案的长期目标是开发一种创新的结构，生物物理和功能性生化框架，以促进我们对HIV剪接机制的理解。 HIV基因组被合成为单个多用电话RNA转录本，该转录物经过调节的剪接以确保mRNA稳态。病毒剪接机械中的畸变会损害HIV复制保真度，在某些情况下会使病毒降低感染力。因此，HIV剪接途径代表了治疗干预的新靶标。剪接决定取决于带有顺式病毒RNA控制元件的反式宿主因子的动态组装/拆卸。但是，确定剪接位点是否会被激活或抑制的基本分子事件仍然难以捉摸。了解一个高度保守的剪接位点如何通过两个反式宿主因子差异调节的物理和功能决定因素提供了定义机制的机会。因此，该提案的直接目标是确定HIV用来选择相互拮抗因子HNRNP A1（A1）和ASF的分子原理，以调节其3'拼接位点A7（SSA7）。此外，A1和ASF通过较差的机制来调节其他保守的HIV剪接位点。因此，这里提出的工作很可能对我们对艾滋病毒剪接的理解具有广泛的影响，并可能导致新的艾滋病毒复制方法。该项目的具体目的是：（1）确定A1和ASF对目标位点选择的贡献，（2）阐明A1和ASF之间相互拮抗作用的生物心理动力学基础，以及（3）研究SSA7调节中合作性的分子起源。