Retrovirus Structure and Assembly with Inositol Phosphates

逆转录病毒结构和用磷酸肌醇组装

• 批准号: 10872754
• 负责人: Robert A Dick
• 金额: $ 12.39万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-05 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10872754
• 关键词: Retrovirus; Structure; Assembly; Inositol; Phosphates

Project Summary Assembly of Gag into an immature virus particle is a critical step in the viral life cycle. After assembly the immature virus particle goes through a process called maturation which is required to produce infectious virus particles. We recently showed that the small cellular molecule, inositol hexakisphosphate (IP6) is required for both assembly and maturation, and that decreasing IP6 levels in cells severely reduces the formation of infectious virus. While we know have a “big picture” view of how IP6 affects HIV, there is still a great deal we don’t understand. This proposal seeks to characterize the mechanism of IP6 induced immature assembly, how IP6 promotes mature assembly, and if these assembly steps can be targeted by antiretrovirals. Furthermore, preliminary work suggests that IP6 is a cofactor for other retroviruses, and so this proposal will test the hypothesis that IP6 as a retroviral cofactor is evolutionarily conserved. Specific Aim 1 is to further characterize how IP6 binds and promotes HIV-1 assembly and maturation. High resolution cryo-EM analysis will be performed on IP6 assembled mature virus-like particles with and without cellular proteins that are known to bind to the viral core in cells. The effect of maturation inhibitors Bevirimat and PF-46396, which bind to the same region as IP6, on IP6 binding and enhanced assembly will be determined using a series of biochemical assays. In addition, this aim will test the hypothesis that IP6 is utilized by other lentiviruses. For example, preliminary data suggest that the Equine Infectious Anemia Virus also employs IP6 for the production of infectious virus. Specific Aim 2 is to determine if and how IP6 is utilized by retroviruses outside of the lentivirus genus. Preliminary data shows that the alpha retrovirus Rous sarcoma virus (RSV), and the delta retrovirus Human T-Cell Leukemia Virus (HTLV) both utilize IP6. We will identify the site of IP6 action on both viruses using biochemical and structural approaches. We will also screen viruses from the other retroviral genera to determine if they also utilize IP6. Specific Aim 3 is to determine if and how IP6 related compounds affect HIV-1 immature assembly, maturation, and mature assembly. Working with a collaborator, we will synthesize and screen IP6-like compounds in assembly and maturation assays. This aim will help to determine if compounds that target the IP6 binding site are a feasible antiretroviral strategy.

项目摘要 将Gag组装成未成熟的病毒颗粒是病毒生命周期中的关键步骤。之后 组装未成熟的病毒颗粒经历了一个称为成熟的过程 产生传染性病毒颗粒所需的物质。我们最近发现，小细胞 分子，六磷酸肌醇(IP6)是组装和成熟所必需的，并且 细胞内IP6水平的降低严重减少了感染性病毒的形成。当我们 我知道对IP6如何影响艾滋病毒有一个“大局”的观点，我们仍然有很多不知道的地方 理解。本研究试图揭示IP6诱导幼稚的机制。 组装，IP6如何促进成熟的组装，以及这些组装步骤是否可以成为 抗逆转录病毒药物。此外，初步工作表明，IP6是其他 因此，这项提议将检验IP6作为逆转录病毒辅助因子的假设 进化上保守的。具体目标1是进一步描述IP6如何结合和促进 HIV-1的组装和成熟。将在IP6上进行高分辨率冷冻-EM分析 组装成熟的病毒样颗粒，含有和不含有已知与之结合的细胞蛋白 细胞中的病毒核心。成熟抑制药贝维利玛和PF-46396对结合力的影响 在IP6结合和增强的组装上与IP6相同的区域将使用 一系列生化分析。此外，这一目标将检验IP6被 其他慢病毒。例如，初步数据表明，马传染性贫血 病毒也使用IP6来生产传染性病毒。具体目标2是确定 以及慢病毒属外的逆转录病毒如何利用IP6。初步数据显示 阿尔法逆转录病毒劳斯肉瘤病毒(RSV)和德尔塔逆转录病毒人类T细胞 白血病病毒(HTLV)都利用IP6。我们将确定IP6在这两种病毒上的作用部位 使用生化和结构方法。我们还将屏蔽来自另一方的病毒 以确定它们是否也利用IP6。具体目标3是确定是否以及如何 IP6相关化合物影响HIV-1的未成熟组装、成熟组装和成熟组装。 与合作者合作，我们将在组装和筛选中合成和筛选类似IP6的化合物 成熟度分析。这一目标将有助于确定靶向IP6结合位点的化合物 是一种可行的抗逆转录病毒策略。

项目成果

Unconventional stabilization of the human T-cell leukemia virus type 1 immature Gag lattice.

人类 T 细胞白血病病毒 1 型未成熟 Gag 晶格的非常规稳定。

• DOI: 10.1101/2023.07.24.548988
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Obr,Martin;Percipalle,Mathias;Chernikova,Darya;Yang,Huixin;Thader,Andreas;Pinke,Gergely;Porley,Dario;Mansky,LouisM;Dick,RobertA;Schur,FlorianKm
• 通讯作者: Schur,FlorianKm

PI(4,5)P2 Clustering and Its Impact on Biological Functions.

PI(4,5)P2 聚类及其对生物功能的影响。

• DOI: 10.1146/annurev-biochem-070920-094827
• 发表时间: 2021
• 期刊: Annual review of biochemistry
• 影响因子: 16.6
• 作者: Wen,Yi;Vogt,VolkerM;Feigenson,GeraldW
• 通讯作者: Feigenson,GeraldW

A Structural Perspective of the Role of IP6 in Immature and Mature Retroviral Assembly.

• DOI: 10.3390/v13091853
• 发表时间: 2021-09-17
• 期刊: Viruses
• 作者: Obr M;Schur FKM;Dick RA
• 通讯作者: Dick RA