CHARACTERIZATION OF HIV-1 PREINTEGRATION COMPLEX ASSEMBLY AND NUCLEAR TRANSPORT

HIV-1 整合前复合体组装和核运输的表征

• 批准号: 7959393
• 负责人: MICHAEL A BELSHAN
• 金额: $ 17.84万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959393
• 关键词: Affinity; Anti-Retroviral Agents; Antiviral Therapy; Area; Biotinylation; Cell Nucleus; Cells; Centrifugation; Complex; Computer Retrieval of Information on Scientific Projects Database; Dependency; Development; Drug resistance; Event; Evolution; Funding; Goals; Grant; HIV; HIV-1; Infection; Institution; Measures; Methods; Nebraska; Proteins; Proteomics; Research; Research Personnel; Resources; Reverse Transcription; Role; Source; Therapeutic; United States National Institutes of Health; inhibitor/antagonist; novel; novel therapeutics; nucleocytoplasmic transport; resistant strain; viral DNA; virology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The evolution of drug resistant strains poses a significant challenge for effective long-term treatment of human immunodeficiency virus type 1 (HIV-1) infection. One counter-measure against resistant strains is treatment with novel therapeutics. Several areas of HIV replication are potential targets for the development of new therapies. The goal of our research is to elucidate the assembly and transport of the HIV preintegration complex (PIC) to aid in the development of novel antiretroviral therapeutics. PICs are large viral DNA complexes formed during early HIV-1 infection of cells that target the viral DNA into the nuclei of cells after reverse transcription. Due to challenges in producing and purifying sufficient quantities of PICs there is limited understanding of their composition, assembly, and mechanism of the transport. Consequently, there are no inhibitors of PICs in development. The objective of our studies is to identify and characterize novel cellular components of HIV-1 PICs. We have undertaken two ambitious strategies to identify PIC-associated cellular proteins. First is a targeted proteomic analysis of PICs partially purified by velocity gradient centrifugation. Second, is a proteomic analysis of PICs affinity purified by specific biotinylation of HIV-1 proteins. Using both methods we have identified several candidate HIV dependency factors (HDFs). Candidate HDFs will be assessed for their interaction with HIV components and their role in HIV replication. Successful completion of these studies will advance the understanding of early events of HIV replication and discover new targets for the development of antiviral therapies.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 耐药毒株的进化对人类免疫缺陷病毒1型(HIV-1)感染的有效长期治疗提出了重大挑战。对抗耐药菌株的一个对策是使用新的治疗方法。艾滋病毒复制的几个领域是开发新疗法的潜在目标。我们研究的目的是阐明HIV前整合复合体(PIC)的组装和运输，以帮助开发新的抗逆转录病毒疗法。PIC是在HIV-1感染细胞的早期形成的大的病毒DNA复合体，在逆转录后将病毒DNA靶向细胞的细胞核。由于在生产和提纯足够数量的PIC方面存在挑战，人们对它们的组成、组装和运输机制的了解有限。因此，目前还没有处于发育阶段的PIC的抑制物。我们研究的目的是确定和表征HIV-1 PIC的新的细胞成分。我们已经采取了两个雄心勃勃的战略来识别PIC相关的细胞蛋白。首先，对速度梯度离心法部分纯化的PIC进行了有针对性的蛋白质组学分析。其次，对通过HIV-1蛋白的特异性生物素化纯化的PICs亲和力进行了蛋白质组学分析。使用这两种方法，我们已经确定了几个候选艾滋病毒依赖因素(HDF)。将评估候选HDF与艾滋病毒组成部分的相互作用及其在艾滋病毒复制中的作用。这些研究的成功完成将促进对艾滋病毒复制早期事件的了解，并为抗病毒疗法的开发发现新的靶点。