HIV-1 capsid assembly intermediates: cellular factors and links to pathogenesis

HIV-1衣壳组装中间体：细胞因素及其与发病机制的联系

基本信息

批准号：
8602634
负责人：
JAISRI R LINGAPPA
金额：
$ 40.85万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-16 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8602634
关键词：
AIDS prevention ATP phosphohydrolase Adaptor Signaling Protein Address Affect Anti-Retroviral Agents Antiviral Agents Area Binding Binding Sites Biochemical CD4 Lymphocyte Count Capsid Cells Characteristics Clinical Collaborations Complex Consensus Cytoplasm Data Disease Outcome Disease Progression Drug Targeting Electrons Environment Enzymes Exhibits Future Gagging Genetic Polymorphism Genomics HIV HIV-1 HIV-2 In Vitro Individual Integration Host Factors Intervention Kinetics Lead Life Cycle Stages Link Macaca Microscopic Modeling Molecular Outcome Pathogenesis Pathogenicity Pathway interactions Patients Pharmaceutical Preparations Production Protein Binding Proteins Publishing RNA RNA Helicase Recruitment Activity Role Site Staging Sum System Testing Variant Viral Viral Load result Virus Virus Assembly base bench to bedside drug candidate drug development gag Gene Products in vivo inhibitor/antagonist insight novel protein protein interaction public health relevance self assembly

项目摘要

DESCRIPTION: In this era in which antiretroviral drugs are being used for both treatment and prevention of AIDS, there is an urgent need for discovery of new drug targets and novel antiretroviral agents. Virus assembly is a complex stage of the HIV-1 life cycle that contains poorly understood drug targets. These targets are understudied, in part because of unresolved controversies in the HIV-1 assembly field. The field has been dominated by the self-assembly model, which is based on the finding that HIV-1 Gag assembles spontaneously in an idealized in vitro system that does not recapitulate the intracellular environment. In contrast, this application is based on a model proposing that the intracellular environment presents barriers to assembly, which the virus overcomes using cellular enzymes. This model is supported by studies demonstrating that during immature capsid assembly, HIV-1 Gag progresses through a stepwise, energy-dependent pathway of assembly intermediates composed of cellular proteins, including two cellular enzymes that facilitate assembly, the ATPase ABCE1 and the RNA helicase DDX6. Thus, while HIV-1 can assemble spontaneously in an idealized in vitro system, studies in cells indicate that HIV-1 capsid assembly is facilitated by at least two host enzymes. Further evidence in favor of the host-catalyzed ABCE1 assembly pathway comes from the discovery by Prosetta Antiviral of novel antiretroviral compounds that target this pathway. This summer Bristol Myers Squibb announced a partnership with Prosetta Antiviral to develop these novel antiretroviral compounds. Despite this breakthrough in the area of drug development, the ABCE1 pathway remains poorly understood. Studies in Aim 1 will test whether ABCE1 acts as an adaptor for recruiting cellular machinery and will also define the Gag-ABCE1 binding site, which appears to involve the highly conserved major homology region of Gag. Aim 2 will help establish a unified model of assembly by determining whether the Gag-containing complex that first associates with HIV-1 genomic RNA, identified by Paul Bieniasz's lab, corresponds to an ABCE1- containing assembly intermediate. Aim 3 will uses viruses generated by Eric Hunter's group to examine whether Gag polymorphisms that arise in vivo can alter viral host interactions, leading to increased virus assembly kinetics, greater virus production, and higher viral loads in infected individuals. Identifying viral-host interactions that impact virus production could lead t strategies for targeting such interactions in the future. In sum, this "bench to bedside" application will advance our understanding of a cellular facilitator of assembly that binds to Gag, help reach a consensus model of HIV-1 capsid packaging, and provide insights into how viral-host interactions in assembly affect pathogenesis.

描述：在抗逆转录病毒药物用于治疗和预防艾滋病的时代，迫切需要发现新的药物靶点和新型抗逆转录病毒药物。病毒组装是 HIV-1 生命周期的一个复杂阶段，其中包含的药物靶点知之甚少。这些目标尚未得到充分研究，部分原因是 HIV-1 组装领域尚未解决的争议。该领域一直以自组装模型为主，该模型基于 HIV-1 Gag 在理想化的体外系统中自发组装的发现，该系统无法重现细胞内环境。相比之下，该应用基于一个模型，该模型提出细胞内环境存在组装障碍，病毒使用细胞酶克服了这些障碍。该模型得到了研究的支持，这些研究表明，在未成熟的衣壳组装过程中，HIV-1 Gag 通过逐步的、能量依赖性的组装中间体途径进行进展，该中间体由细胞蛋白组成，包括两种促进组装的细胞酶，ATPase ABCE1 和 RNA 解旋酶 DDX6 。因此，虽然 HIV-1 可以在理想的体外系统中自发组装，但细胞研究表明，至少两种宿主酶可促进 HIV-1 衣壳组装。支持宿主催化 ABCE1 组装途径的进一步证据来自 Prosetta Antiviral 针对该途径的新型抗逆转录病毒化合物的发现。今年夏天，百时美施贵宝宣布与 Prosetta Antiviral 合作开发这些新型抗逆转录病毒化合物。尽管药物开发领域取得了这一突破，但 ABCE1 通路仍然知之甚少。目标 1 中的研究将测试 ABCE1 是否充当招募细胞机器的接头，并将定义 Gag-ABCE1 结合位点，该位点似乎涉及 Gag 高度保守的主要同源区域。目标 2 将通过确定 Paul Bieniasz 实验室鉴定的首先与 HIV-1 基因组 RNA 结合的包含 Gag 的复合物是否对应于包含 ABCE1 的装配中间体，来帮助建立统一的装配模型。 Aim 3将使用Eric Hunter小组产生的病毒来检查体内出现的Gag多态性是否可以改变病毒宿主相互作用，从而导致感染个体中病毒装配动力学增强、病毒产量增加和病毒载量增加。识别影响病毒产生的病毒与宿主的相互作用可能会导致未来针对此类相互作用的策略。总之，这种“从工作台到床边”的应用将增进我们对与 Gag 结合的细胞组装促进剂的理解，有助于达成 HIV-1 衣壳包装的共识模型，并提供有关装配中病毒与宿主相互作用如何影响发病机制的见解。