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：
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
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/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

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：在这个时代，抗逆转录病毒药物既用于治疗和预防艾滋病，迫切需要发现新药物靶标和新型抗逆转录病毒药物。病毒组装是HIV-1生命周期的复杂阶段，其中包含知识渊博的药物靶标。这些目标被研究了，部分原因是HIV-1组装领域未解决的争议。该领域由自组装模型主导，该模型基于以下发现：HIV-1插入在理想化的体外系统中自发地组装，该系统不会概括细胞内环境。相反，该应用是基于一个模型，该模型表明细胞内环境为组装壁垒提供了障碍，该障碍使用细胞酶克服了该病毒。该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通过由细胞蛋白组成的组装中间体的逐步，能量依赖性途径进行，包括两个促进组装的细胞酶，促进ATPase ABCE1和RNA Helicase ddx66 。因此，尽管HIV-1可以在理想化的体外系统中自发地组装，但细胞的研究表明HIV-1 CAPSID组装由至少两种宿主酶促进。支持宿主催化的ABCE1组装途径的进一步证据来自Prosetta抗病毒对靶向该途径的新型抗逆转录病毒化合物的发现。今年夏天，布里斯托尔·迈尔斯（Bristol Myers Squibb）宣布与Prosetta抗病毒建立合作伙伴关系，以开发这些新颖的抗逆转录病毒化合物。尽管在药物开发领域取得了突破，但ABCE1途径仍然知之甚少。 AIM 1中的研究将测试ABCE1是否充当募集细胞机械的适配器，还将定义GAG-ABCE1结合位点，该位点似乎涉及高度保守的GAG主要同源性区域。 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首次关联的含GAG的复合物来帮助建立统一的组装模型，该复合物是否对应于含有ABCE1的含有ABCE1的组装中间体。 AIM 3将使用埃里克·亨特（Eric Hunter）小组生成的病毒来检查体内产生的插孔多态性是否可以改变病毒宿主的相互作用，从而导致病毒组装动力学，更多的病毒产生和更高的感染个体病毒负荷。识别影响病毒生产的病毒宿主相互作用可能会导致未来针对此类相互作用的策略。总而言之，这种“板凳至床头”的应用将提高我们对与插科打结合的组件的蜂窝促进者的理解有助于达到HIV-1 CAPSID包装的共识模型，并提供有关组装中病毒宿主相互作用如何影响发病机理的见解。