CONTRIBUTION OF CELLULAR FACTOR TO HIV-1 ASSEMBLY

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 A.具体目标 全球艾滋病毒疫情继续扩大，超出了以前的预测，已成为人类历史上最致命的流行病之一。非裔美国妇女感染艾滋病毒的高流行率表明，需要制定新的医疗干预措施，以消除妇女在艾滋病毒/艾滋病方面的健康差距。对目前批准的药物具有耐药性的艾滋病毒-1分离株的出现和传播使发现具有新机制和目标的新型抗艾滋病毒药物成为高度优先的研究重点。HIV-1 Gag蛋白指导着高度有序的颗粒组装和释放过程。目前正在确定艾滋病毒-1复制周期的这些后期阶段所涉及的不同步骤，但我们的知识仍需填补重大空白。最近，通过酵母双杂交筛选人的cDNA文库，我们发现了一个新的Gag结合伙伴--丝氨酸A。丝氨酸A是一种非肌肉肌动蛋白结合蛋白，在将皮质细丝交联成动态三维结构中起着重要作用。Flna与不同的细胞蛋白相互作用，是蛋白质运输、信号转导以及细胞-细胞和/或细胞-基质连接所需的多功能支架。在HIV-1颗粒组装和释放过程中发现Flna-Gag相互作用，表明Flna通过调节肌动蛋白细胞骨架重塑促进HIV-1 Gag转运到质膜。这一RCMI试点项目的总体目标是确定FLNA-GAG相互作用的分子基础及其生物学意义。我们的研究将提供有关逆转录病毒-宿主相互作用的重要新信息，并将通过发现和开发新的组装抑制剂来影响抗HIV治疗。这一建议将通过在三个综合具体目标内组织的一系列实验来完成。 具体目标1：确定FLNA-GAG相互作用的分子基础。 具体目标2：确定Flna调控的HIV-1 Gag贩运的机制。 具体目标3：确定Flna在人原代CD4T细胞和巨噬细胞中的作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. A. Specific Aims The global HIV epidemic continues to expand exceeding previous predictions and has become one of the deadliest epidemics in human history. The high prevalence of HIV infection in the African-American women points to the need to develop the new medical interventions toward eliminating women's health disparities in HIV/AIDS. The emergence and transmission of HIV-1 isolates resistant to currently approved drugs makes the discovery of novel anti-HIV drugs with new mechanisms and targets a high research priority. HIV-1 Gag protein directs the highly ordered process of particle assembly and release. Distinct steps involved in these late stages of the HIV-1 replication cycle are being defined, yet significant gaps still need to be filled in our knowledge. Recently, by yeast two-hybrid screening of a human cDNA library, we identified a novel Gag-binding partner, filamin A. Filamin A (FLNa) is a non-muscle actin binding protein that plays an important role in cross-linking cortical filaments into a dynamic three-dimensional structure. FLNa interacts with different cellular proteins, and serves as a versatile scaffold required for protein trafficking, signaling transduction, and cell-cell and/or cell-matrix connections. The discovery of the FLNa-Gag interaction in a productive manner in HIV-1 particle assembly and release suggests that FLNa facilitates HIV-1 Gag trafficking to the plasma membrane by regulating the actin cytoskeleton remodeling. The overall goal of this RCMI pilot project is to define the molecular basis of the FLNa-Gag interaction and its biological significance. Our studies will provide important new information regarding retrovirus-host interactions, and will impact anti-HIV therapy by discovering and developing novel assembly inhibitors. This proposal will be accomplished in a series of experiments organized within three integrated specific aims. Specific Aim 1: To define the molecular basis of the FLNa-Gag interaction. Specific Aim 2: To define the mechanism of FLNa-regulated HIV-1 Gag trafficking. Specific Aim 3: To define the role of FLNa in human primary CD4+ T cells and macrophages.