Project 5: Structural investigation of lentiviral DNA integration

P5。摘要 逆转录病毒复制依赖于将逆转录病毒DNA整合到宿主细胞染色体中。这 这一过程是由整合酶(IN)在稳定的突触复合体(称为肠小体)的背景下催化的。这个 Intasome是INSTI的靶标，INSTI是目前唯一的HIV IN拮抗剂 被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内切酶进行了鉴定， 共同阐明负责整合的保守的内体核心组件(CIC)。我们现在有了 建立了第一个基于Maedi-Visna病毒(MVV)的慢病毒肠炎模型，该模型提供了详细的 在IN中没有增溶或过度激活突变的慢病毒整合的结构研究。这个 切里帕诺夫实验室用单粒子冷冻EM对MVV内切酶进行的初步研究表明，它 比其非慢病毒对应物大得多，由IN的十六聚体(四聚体)组成。在……里面 我们利用新的系统来研究慢病毒DNA整合到染色质中。 以及宿主细胞因子LEDGF/p75在这一过程中的作用。作为该项目的翻译部分，我们 将为INSTI的结构和耐药机制研究开发一种慢病毒肠酶模型。

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.