Mechanisms and Consequences of Reverse Transcription in HIV-1 Cores

HIV-1 核心逆转录的机制和后果

• 批准号: 10454310
• 负责人: Christopher R Aiken
• 金额: $ 68.08万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 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/10454310
• 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; insight; mutant; nucleic acid binding protein; particle; scaffold; therapeutic target; therapy resistant; uptake; viral DNA; viral RNA; viral genomics

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.

总结 HIV-1感染导致T辅助细胞的破坏，导致免疫缺陷和免疫缺陷。 被称为艾滋病的疾病。大多数目前可用的抗逆转录病毒药物靶向病毒逆转 转录酶、整合酶和蛋白酶。这些药物有效，但不能治愈， 治疗必须严格和持久地坚持。此外，HIV-1不断进化， 并获得突变使其对治疗产生抗性。因此，需要进行持续的研究 来开发针对新的病毒和细胞靶点的药物。由于其在感染中的多重作用， HIV-1衣壳是一个有吸引力的治疗靶点， 结构在临床试验中显示出有希望的结果。衣壳是一个组装的晶格 由单个病毒蛋白组成，在病毒基因组和相关蛋白周围形成外壳 并且对于HIV-1感染的关键早期步骤有效逆转录是必需的。 对衣壳稳定性的扰动导致感染失败，但感染失败的机制仍然存在。 衣壳确保有效的逆转录是未知的。该项目将填补这一空白， 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颗粒中纯化的天然病毒核心，我们将采用尖端的 生物物理技术与计算和分子病毒学方法一起， 精确定义了HIV-1逆转录中病毒衣壳的作用。该项目将 完成以下目标：（1）确定逆转录的生物化学要求 在HIV-1核心中;（2）在逆转录过程中定义核心中的结构转变;（3） 确定逆转录期间衣壳孔在核苷酸摄取中的作用;和（4） 精确地定义在特定阶段的核心内的蛋白质-核酸相互作用， 逆转录总的来说，这个项目将产生一个更好的理解衣壳功能 在HIV-1感染中，从而为一类新兴的抗病毒药物的作用机制提供信息。 毒品 相关性 该项目的结果将是更好地了解病毒衣壳的功能， 是HIV-1感染的关键。这些信息将为capture-targeting的发展提供信息 抗病毒药物和改善艾滋病毒感染者的治疗，从而改善公众健康 减少HIV-1的传播。