Project 5: Structural investigation of lentiviral DNA integration

项目5：慢病毒DNA整合的结构研究

• 批准号: 10219103
• 负责人: Peter Cherepanov
• 金额: $ 30.2万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-27 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10219103
• 关键词: AIDS/HIV problem; Active Sites; Amino Acid Sequence; Architecture; Area; Binding; Cells; Chromatin; Chromatin Structure; Chromosomes; Clinical; Code; Complex; Core Assembly; Cryoelectron Microscopy; Crystallization; DNA Integration; Drug resistance; Epigenetic Process; Genes; Genetic Transcription; HIV; HIV Integrase; HIV-1; HIV-1 integrase; Integrase; Integration Host Factors; Investigation; Laboratories; Length; Lentivirus; Modeling; Mutagenesis; Mutation; Nature; Orthologous Gene; Play; Process; Proteins; Refractory; Resolution; Role; Site; Spumavirus; Structural Models; Structure; Synapses; System; Testing; Virus; Visna-maedi virus; Visualization; base; bone; inhibitor/antagonist; lentiviral integration; particle; preference; prototype; small molecule; success; transcriptional coactivator p75; viral DNA; viral resistance

P5. Abstract Retroviral replication depends on integration of reverse transcribed viral DNA into a host cell chromosome. This process is catalyzed by integrase (IN) in the context of a stable synaptic complex, known as the intasome. The intasome is the target for IN strand transfer inhibitors (INSTIs), the only class of HIV IN antagonists currently approved to treat HIV/AIDS. Recent years saw characterization of the intasomes from several retroviral species, collectively elucidating the conserved intasomal core assembly (CIC) responsible for integration. We have now established the first lentiviral intasome model based on maedi-visna virus (MVV), which affords detailed structural studies of lentiviral integration in the absence of solubilizing or hyperactivating mutations in IN. The preliminary study of the MVV intasome by single particle cryo-EM in the Cherepanov laboratory revealed that it is much larger than its non-lentiviral counterparts, comprising a hexadecamer (tetramer-of-tetramers) of IN. In the proposed project we take advantage of the new system to study lentiviral DNA integration into chromatin and the role of the host cell factor LEDGF/p75 in this process. As a translational component of this project, we will develop a lentiviral intasome model for structural studies of INSTIs and the mechanisms of drug resistance.

P5。抽象的 逆转录病毒复制依赖于逆转录病毒DNA整合到宿主细胞染色体中。这 该过程由整合酶（IN）在稳定的突触复合物（称为整合体）的背景下催化。这 intasome 是 IN 链转移抑制剂 (INSTI) 的靶标，INSTI 是目前唯一一类 HIV IN 拮抗剂 批准用于治疗艾滋病毒/艾滋病。近年来，人们对几种逆转录病毒物种的嵌体进行了表征， 共同阐明了负责整合的保守的内体核心组件（CIC）。我们现在有 建立了第一个基于maedi-visna病毒（MVV）的慢病毒嵌体模型，该模型提供了详细的 在IN中不存在溶解或过度激活突变的情况下慢病毒整合的结构研究。这 Cherepanov 实验室通过单粒子冷冻电镜对 MVV 整合体进行的初步研究表明， 比其非慢病毒对应物大得多，由 IN 的十六聚体（四聚体的四聚体）组成。在 在拟议的项目中，我们利用新系统来研究慢病毒 DNA 整合到染色质中 以及宿主细胞因子 LEDGF/p75 在此过程中的作用。作为该项目的翻译组成部分，我们 将开发慢病毒嵌体模型，用于 INSTI 的结构研究和耐药机制。