Novel host proteins in the HIV-1 preintegration complexes

HIV-1预整合复合物中的新型宿主蛋白

• 批准号: 8410611
• 负责人: Li Wu
• 金额: $ 19.06万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-20 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8410611
• 关键词: Address; Affect; Anti-HIV Agents; Binding Proteins; Biological Assay; CD4 Positive T Lymphocytes; Cell Survival; Cells; Complementary DNA; Complex; DNA; DNA Repair; Drug Delivery Systems; Drug resistance; Funding Mechanisms; Goals; HIV; HIV Infections; HIV-1; Human; Individual; Infection; Integrase; Knowledge; Length; Lentivirus Vector; Life; Measures; Messenger RNA; Nature; Nuclear Import; Nuclear Protein; Play; Process; Proteins; RNA Splicing; Relative (related person); Role; Testing; Therapeutic; Transcriptional Regulation; Viral; Viral Proteins; Virus Diseases; antiretroviral therapy; base; cofactor; design; insight; mRNA Expression; novel; novel strategies; novel therapeutics; prevent; small hairpin RNA

DESCRIPTION (provided by applicant): The HIV-1 pre-integration complex (PIC) that comprises of HIV-1 full-length DNA, viral proteins, and host proteins is essential for viral integration and replication. Previous studies have identified and characterized several human proteins that interact with HIV-1 integrase and play important roles in viral infection. However, the technical limitations in acquiring sufficient amounts of catalytically active PICs has hindered a comprehensive identification of host proteins associated with the HIV-1 PICs. We recently developed a novel approach and identified 18 new human proteins that are specifically associated with the HIV-1 PICs isolated from infected CD4+ T cells. Our preliminary study suggested that one of these proteins, non-POU domain-containing octamer-binding protein (NonO), is required for efficient HIV-1 infection in CD4+ T cells. However, it is unclear whether the other 17 PIC- associated host proteins that we identified can affect HIV-1 infection. NonO is a multifunctional nuclear protein involved in transcription regulation, mRNA splicing, and DNA repair in the cell, but the mechanism by which NonO affects HIV-1 infection is not known. We seek to address these two significant questions in this R21 proposal. We hypothesize that host proteins (such as NonO) specifically associated with catalytically active PICs in HIV-1-infected cells play a role in viral integration and infection. The following two specific aims are designed o test this hypothesis: Aim 1. To determine the role of novel PIC-associated host proteins in HIV-1 infection. In addition to NonO, we may identify other PIC-associated host proteins among these 18 candidates that can significantly affect HIV-1 infection in Aim 1. We will select 2-3 protein candidates that have the most significant effect on viral infection to further study their mechanisms of action. Aim 2. To investigate the mechanisms by which PIC-associated host proteins affect HIV-1 infection. We will first investigate the mechanism by which NonO affects HIV-1 infection, and we will then perform similar studies to examine other protein candidates identified in Aim 1. The long-term goal of this project is to understand the function and mechanism of PIC-associated host proteins in HIV-1 integration or infection and to develop novel therapeutic strategies. An emerging anti-HIV therapeutic strategy is to block the interactions between HIV-1 integrase and its critical cellular cofactors. Our proposed study has the potential to identify novel drug targets to block HIV-1 integration and infection. PUBLIC HEALTH RELEVANCE: There are over 33 million people living with HIV worldwide and approximately 56,300 new HIV infections each year in the US. Although antiretroviral therapy can control HIV-1 infection in infected individuals, new anti-HIV drug targets are needed to increase efficacy and prevent drug resistance. This project aims to identify and characterize novel host proteins that are essential for the HIV integration process. The findings from the proposed studies will help identify novel drug targets for blocking HIV integration and infection.

描述（由申请人提供）：HIV-1前整合复合物（PIC）包括HIV-1全长DNA，病毒蛋白和宿主蛋白，对于病毒整合和复制至关重要。先前的研究已经确定并表征了几种与HIV-1整合酶相互作用并在病毒感染中起重要作用的人类蛋白质。但是，获取足够数量的催化活性图片的技术限制已阻碍 与HIV-1图片相关的宿主蛋白的全面鉴定。我们最近开发了一种新颖的方法，并确定了18种新的人蛋白，这些蛋白质与从感染的CD4+ T细胞中分离出的HIV-1 PIC特别相关。我们的初步研究表明，在CD4+ T细胞中有效HIV-1感染需要这些蛋白质之一，即非POU结构域的八聚体结合蛋白（NONO）。但是，尚不清楚我们确定的其他17个相关的宿主蛋白是否会影响HIV-1感染。 NONO是一种参与细胞中转录调节，mRNA剪接和DNA修复的多功能核蛋白，但是Nono影响HIV-1感染的机制尚不清楚。我们试图在此R21提案中解决这两个重要问题。我们假设与HIV-1感染细胞中催化活性图片特别相关的宿主蛋白（例如NONO）在病毒整合和感染中起作用。设计以下两个特定目标o检验以下假设：目标1。确定新型PIC相关宿主蛋白在HIV-1感染中的作用。除NONO外，我们还可以在这18个候选者中鉴定出其他与PIC相关的宿主蛋白，这些候选者可能会在AIM 1中显着影响HIV-1感染。我们将选择对病毒感染具有最显着影响的2-3个蛋白质候选者，以进一步研究其作用机理。目的2。研究与PIC相关宿主蛋白影响HIV-1感染的机制。我们将首先研究NONO影响HIV-1感染的机制，然后我们将进行类似的研究，以检查AIM 1中鉴定的其他蛋白质候选物。该项目的长期目标是了解HIV-1整合或感染中PIC相关宿主蛋白的功能和机制，并开发新的治疗策略。新兴的抗HIV治疗策略是阻止HIV-1积分酶与其关键细胞辅助因子之间的相互作用。我们提出的研究有潜力鉴定新的药物靶标，以阻止HIV-1整合和感染。 公共卫生相关性：全世界有超过3,300万人患HIV，在美国每年大约有56,300例新的艾滋病毒感染。尽管抗逆转录病毒疗法可以控制感染个体的HIV-1感染，但仍需要新的抗HIV药物靶标以提高功效并防止耐药性。该项目旨在识别和表征对HIV整合过程至关重要的新型宿主蛋白。拟议研究的发现将有助于确定阻止HIV整合和感染的新型药物靶标。