Mechanisms and Consequences of Reverse Transcription in HIV-1 Cores

HIV-1 核心逆转录的机制和后果

基本信息

批准号：
10337946
负责人：
Christopher R Aiken
金额：
$ 71.73万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
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：
2021-07-20 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10337946
关键词：
Acquired Immunodeficiency Syndrome Amino Acid Substitution Anti-Retroviral Agents Antiviral Agents Atomic Force Microscopy Binding Binding Sites Biochemical Biological Assay Biological Metamorphosis Capsid Cells Characteristics Clinical Trials Complex Computer Models DNA biosynthesis Development Disease Drug Targeting Ensure Enzymes Exhibits Genetic Transcription Goals HIV HIV-1 Helper-Inducer T-Lymphocyte High-Throughput Nucleotide Sequencing Immunologic Deficiency Syndromes Immunoprecipitation In Vitro Infection Integrase Length Maps Medicine Molecular Molecular Virology Mutation Negative Staining Nucleic Acids Nucleoproteins Nucleotides Pattern Penetration Peptide Hydrolases Perilla Persons Pharmaceutical Preparations Phytic Acid Proteins Public Health RNA Binding RNA-Directed DNA Polymerase Radiolabeled Reaction Research Reverse Transcription Role Series Structure Study models System Technology Testing Transmission Electron Microscopy Viral Viral Genome Viral Physiology Viral Proteins Virus Diseases Work biophysical techniques cellular targeting crosslink crosslinking and immunoprecipitation sequencing deep sequencing ds-DNA experimental study genomic RNA improved inhibitor/antagonist insight mutant nucleic acid binding protein particle scaffold therapeutic target therapy resistant uptake viral DNA viral RNA viral genomics

项目摘要

Summary HIV-1 infection results in destruction of T helper cells, leading to immunodeficiency and the disease known as AIDS. Most currently available antiretroviral drugs target the viral reverse transcriptase, integrase, and protease enzymes. These medicines are effective but not curative, and therapy must be adhered to rigorously and permanently. Moreover, HIV-1 constantly evolves and acquires mutations rendering it resistant to therapies. Therefore, ongoing research is needed to develop drugs against new viral and cellular targets. Owing to its multiple functions in infection, the HIV-1 capsid represents an attractive therapeutic target, and potent inhibitors targeting this structure have shown promising results in clinical trials. The capsid, which is an assembled lattice composed of a single viral protein, forms a shell around the viral genome and associated proteins and is essential for efficient reverse transcription, a key early step in HIV-1 infection. Perturbations to the stability of the capsid result in abortive infection, yet the mechanism by which the capsid ensures efficient reverse transcription is unknown. This project will fill this gap by defining the effects of capsid perturbations on HIV-1 reverse transcription in vitro, and vice-versa. Using native viral cores purified from infectious HIV-1 particles, we will employ cutting-edge biophysical techniques together with computational and molecular virology approaches to precisely define the role of the viral capsid in HIV-1 reverse transcription. The project will accomplish the following goals: (1) define the biochemical requirements for reverse transcription in HIV-1 cores; (2) define the structural transitions in the core during reverse transcription; (3) determine the role of capsid pores in nucleotide uptake during reverse transcription; and (4) precisely define the protein-nucleic acid interactions within the core during specific stages of reverse transcription. Collectively, this project will yield a greater understanding of capsid function in HIV-1 infection, thereby informing the mechanism of action of an emerging class of antiviral drugs. Relevance The results of this project will be a greater understanding of the functions of the viral capsid that are critical for HIV-1 infection. This information will inform the development of capsid-targeting antiviral drugs and improve the treatment of HIV-infected persons, thereby improving public health and reducing the spread of HIV-1.

概括 HIV-1感染导致T辅助细胞破坏，导致免疫缺陷和被称为艾滋病的疾病。目前最可用的抗逆转录病毒药物针对病毒反向转录酶，整合酶和蛋白酶酶。这些药物有效但不能治愈，并且必须严格和永久地遵守治疗。而且，HIV-1不断发展并获得对疗法具有抵抗力的突变。因此，需要进行持续的研究开发针对新病毒和细胞靶标的药物。由于其在感染中的多个功能， HIV-1衣壳代表了一个有吸引力的治疗靶标，并且针对此目标的有效抑制剂结构在临床试验中显示出令人鼓舞的结果。衣壳，这是一个组装的晶格由单个病毒蛋白组成，在病毒基因组和相关蛋白周围形成壳对于有效的逆转录至关重要，这是HIV-1感染中的关键早期步骤。对衣壳的稳定性的扰动导致流产感染，但其机制衣壳确保有效的逆转录未知。这个项目将通过定义衣壳扰动对体外HIV-1逆转录的影响，反之亦然。使用从感染性HIV-1颗粒纯化的天然病毒核心，我们将采用尖端生物物理技术以及计算和分子病毒学方法精确地定义了病毒包囊在HIV-1逆转录中的作用。该项目将完成以下目标：（1）定义逆转录的生化要求在HIV-1核心；（2）定义逆转录过程中核心中的结构跃迁；（3）确定在逆转录过程中胶囊孔在核苷酸摄取中的作用；（4）精确定义核心内蛋白核酸相互作用在特定阶段逆转录。总体而言，该项目将对CAPSID功能有更深入的了解在HIV-1感染中，从而告知新兴类抗病毒药的作用机理毒品。关联该项目的结果将是对病毒式衣壳功能的更深入的了解对于HIV-1感染至关重要。此信息将为capsid靶向的开发提供信息抗病毒药物并改善对艾滋病毒感染者的治疗，从而改善公共卫生并减少HIV-1的传播。