Novel host proteins in the HIV-1 preintegration complexes

性状（由申请方提供）：HIV-1整合前复合物（PIC）由HIV-1全长DNA、病毒蛋白和宿主蛋白组成，对病毒整合和复制至关重要。先前的研究已经鉴定和表征了几种与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。然而，在获得足够量的催化活性PIC方面的技术限制阻碍了催化剂的应用。 全面鉴定与HIV-1 PIC相关的宿主蛋白。我们最近开发了一种新的方法，并确定了18种新的人类蛋白质，这些蛋白质与从感染的CD 4 + T细胞中分离的HIV-1 PIC特异性相关。我们的初步研究表明，这些蛋白质之一，非POU结构域的八聚体结合蛋白（NonO），需要有效的HIV-1感染的CD 4 + T细胞。然而，目前还不清楚我们鉴定的其他17种PIC相关宿主蛋白是否可以影响HIV-1感染。NonO是一种多功能核蛋白，参与细胞中的转录调节、mRNA剪接和DNA修复，但NonO影响HIV-1感染的机制尚不清楚。我们力求在21国责任的建议中解决这两个重要问题。我们推测，宿主蛋白（如NonO）与HIV-1感染细胞中的催化活性PIC特异性相关，在病毒整合和感染中发挥作用。以下两个具体目标是为了检验这一假设：目标1。确定新型PIC相关宿主蛋白在HIV-1感染中的作用。除了NonO之外，我们还可以在这18种候选蛋白中鉴定出其他PIC相关的宿主蛋白，这些蛋白可以显著影响Aim 1中的HIV-1感染。我们将选择2-3种对病毒感染具有最显著影响的候选蛋白，以进一步研究其作用机制。目标二。研究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，以检查目标1中确定的其他候选蛋白。该项目的长期目标是了解PIC相关宿主蛋白在HIV-1整合或感染中的功能和机制，并开发新的治疗策略。一种新兴的抗HIV治疗策略是阻断HIV-1整合酶与其关键细胞辅因子之间的相互作用。我们提出的研究有可能确定新的药物靶点，以阻止HIV-1整合和感染。

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.