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

• 批准号: 9039524
• 负责人: JAISRI R LINGAPPA
• 金额: $ 43.5万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 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/9039524
• 关键词: ABCE1 gene; AIDS prevention; ATP phosphohydrolase; Adaptor Signaling Protein; Address; Affect; Anti-Retroviral Agents; Antiviral Agents; Area; Binding; Binding Proteins; 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; Genetic Polymorphism; 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; Proteins; Publishing; 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; genomic RNA; in vivo; inhibitor/antagonist; insight; new therapeutic target; novel; protein protein interaction; public health relevance; self assembly

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：在这个抗逆转录病毒药物被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的抗逆转录病毒药物。病毒组装是HIV-1生命周期的一个复杂阶段，其中包含鲜为人知的药物靶点。对这些靶点的研究不足，部分原因是艾滋病毒-1组装领域尚未解决的争议。该领域一直被自组装模型所主导，该模型基于这样的发现，即HIV-1 Gag在一个理想化的体外系统中自发组装，而不是概括细胞内环境。相比之下，这一应用是基于一个模型，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一障碍。这一模型得到了研究的支持，该研究表明，在未成熟的衣壳组装过程中，HIV-1 Gag通过一条循序渐进的、依赖能量的途径进行组装，中间产物由细胞蛋白组成，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，但细胞中的研究表明，至少有两种宿主酶促进了HIV-1衣壳的组装。支持宿主催化的ABCE1组装途径的进一步证据来自Prosetta抗病毒公司发现了针对这一途径的新型抗逆转录病毒化合物。今年夏天，百时美施贵宝宣布与Prosetta AntiVirus合作开发这些新型抗逆转录病毒化合物。尽管在药物开发领域取得了这一突破，但ABCE1途径仍然知之甚少。AIM 1的研究将测试ABCE1是否作为招募细胞机械的适配器，并定义GAG-ABCE1结合位点，这似乎涉及GAG的高度保守的主要同源区。AIM 2将通过确定由Paul Bieniasz的实验室鉴定的首先与HIV-1基因组RNA结合的含有Gag的复合体是否对应于含有ABCE1的组装中间体，来帮助建立一个统一的组装模型。AIM 3将使用Eric Hunter的团队产生的病毒来检查体内出现的Gag多态是否可以改变病毒宿主的相互作用，从而导致感染个体的病毒组装动力学增加、病毒产量增加和病毒载量增加。识别影响病毒产生的病毒-宿主相互作用可能导致未来针对此类相互作用的策略。总而言之，这种“工作台到床边”应用程序将促进我们对绑定到GAG的组装的蜂窝促进器的理解， 帮助达成HIV-1衣壳包装的共识模型，并提供对组装中病毒与宿主相互作用如何影响发病机制的见解。