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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 抗药性菌株的演变对有效的长期治疗人类免疫缺陷病毒1型（HIV-1）感染构成了重大挑战。对抗性菌株的一种对立是对新型治疗药的治疗。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前整合络合物（PIC）的组装和运输，以帮助开发新型的抗逆转录病毒疗法。图片是在逆转录后将病毒DNA靶向细胞核的细胞的早期HIV-1感染中形成的大型病毒DNA复合物。由于在产生和净化足够数量的图片方面面临挑战，因此对它们的组成，组装和运输机制的理解有限。因此，没有图片的抑制剂在开发中。我们研究的目的是识别和表征HIV-1图片的新细胞成分。我们采取了两种雄心勃勃的策略来鉴定与PIC相关的细胞蛋白。首先是对图片的靶向蛋白质组学分析，该图片通过速度梯度离心部分纯化。其次，是通过HIV-1蛋白的特异性生物素化纯化的图片亲和力的蛋白质组学分析。使用这两种方法，我们都确定了几个候选HIV依赖性因素（HDF）。将评估候选HDF与HIV成分的相互作用及其在HIV复制中的作用。这些研究的成功完成将提高人们对艾滋病毒复制早期事件的理解，并发现开发抗病毒疗法的新目标。