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: 7609844
• 负责人: MICHAEL A BELSHAN
• 金额: $ 18.71万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609844
• 关键词: Anti-Retroviral Agents; Architecture; Biological Assay; Cell Nucleus; Cells; Class; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Funding; Grant; HIV; HIV-1; In Vitro; Institution; Longevity; Methods; Nuclear Import; Nucleic Acids; Nucleoproteins; Proteins; Research; Research Personnel; Resources; Reverse Transcription; Source; Therapeutic; United States National Institutes of Health; Viral; gel electrophoresis; insight; nucleocytoplasmic transport; size; tandem mass spectrometry; viral DNA

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 experimental focus of this proposal is to investigate the composition, assembly and transport of the human immunodeficiency virus (HIV) preintegration complex (PIC). The PIC is a large nucleoprotein complex that facilitates transport of the viral DNA into the nucleus of cells upon completion of reverse transcription. The viral components of the PIC have been characterized, but the size of the PIC suggests it contains a large amount of cellular factors, perhaps both protein and nucleic acid. Structural studies of PICs are hampered by several factors, including: 1) Low quantity in the cell; 2) a relatively short life-span; and 3) difficulty of purification. We propose to develop methods to produce and purify a sufficient quantity of PICs to proteomically analyze PIC composition and characterize nuclear transport. Composition will be analyzed by gel electrophoresis and tandem mass spectrometry. Nuclear transport will be assessed by an in vitro nuclear import and integration assay. This data will give insight into the architecture and cellular transport of the PIC, the first step in developing a new class of anti-retroviral therapeutics.

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。 该提案的实验重点是研究人类免疫缺陷病毒（HIV）前整合复合物（PIC）的组成，组装和运输。 PIC是一种大型核蛋白复合物，可在逆转录完成后促进病毒DNA传输到细胞核中。 PIC的病毒成分已经表征了，但是PIC的大小表明它包含大量的细胞因子，也许均可能蛋白质和核酸。图片的结构研究受到几个因素的阻碍，包括：1）细胞中的低数量； 2）相对较短的寿命； 3）纯化难度。我们建议开发方法来生产和净化足够数量的图片，以分析PIC组成并表征核转运。组成将通过凝胶电泳和串联质谱法分析。核运输将通过体外核进口和整合测定法进行评估。这些数据将洞悉PIC的结构和细胞运输，这是开发新的抗逆转录病毒疗法的第一步。