Cellular Proteins Involved in Trafficking of HIV-1

参与 HIV-1 贩运的细胞蛋白

• 批准号: 7618181
• 负责人: Carol A Carter
• 金额: $ 33.93万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-15 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7618181
• 关键词: Agonist; Antibodies; Antiviral Agents; Antiviral Therapy; Binding; Biochemical; Biological; Birds; Calcium; Calcium Channel; Calcium Signaling; Cell Line; Cell membrane; Cell physiology; Cells; Coupled; Cytosol; Development; Environment; Event; Exocytosis; Family; Family member; Fluorescence; Gagging; Genes; Genetic; Goals; HIV; HIV-1; Hybrids; ITPR1 gene; Inositol; International; Intervention; Investigation; Knock-out; Laboratories; Lead; Ligands; Link; Mediating; Microinjections; Multivesicular Body; Mutate; Mutation; Pharmaceutical Preparations; Polyproteins; Predisposition; Process; Protein Biochemistry; Protein Family; Proteins; Recruitment Activity; Reporting; Research Personnel; Resistance; Retroviridae; Role; Rous sarcoma virus; Signal Transduction; Site; Small Interfering RNA; TSG101 gene; Testing; Ubiquitin; Vaccines; Variant; Viral; Virus; Virus Assembly; Virus-like particle; base; extracellular; gag Gene Products; inhibitor/antagonist; insight; late endosome; novel; pandemic disease; particle; protein function; receptor; trafficking; transmission process; tripolyphosphate; ubiquitin ligase

DESCRIPTION (provided by applicant): The human immunodeficiency virus type 1 (HIV-1) is the causative agent of a national and international pandemic. No vaccine is currently available and viral variants resistant to current drug-based antiviral therapies are emerging world-wide. These facts make it imperative to identify new targets and strategies for development of anti-viral drugs. The gag gene of HIV and other retroviruses encode the viral structural precursor polyprotein, Gag, and is sufficient for assembly and release of virus-like particles. Many cellular proteins contribute to this process and represent potential novel targets. Our preliminary studies suggest that the inositol (1,4,5)-triphosphate receptor (IP3R) is involved in trafficking of HIV-1 Gag and the related avian Rous Sarcoma Virus (RSV) Gag to release sites on the plasma membrane. The IP3R protein has not been previously linked to retroviral trafficking. IP3R forms a ligand-gated calcium channel that regulates the release of intracellular calcium stores. Calcium release regulates many cellular processes, including intracellular trafficking and exocytosis. Inhibitors of IP3R interfere with Gag trafficking. Susceptibility to IP3R inhibitors requires the Late domains in the HIV and RSV Gag proteins, which are regions that bind Tsg101 and Nedd4, respectively. Tsg101 and Nedd4 are cellular proteins that facilitate trafficking of HIV and RSV Gag, respectively, to sites on the plasma membrane where Gag assembles into virus-like particles (VLPs) and buds into the extracellular environment. The goals of the proposed studies are (AIM 1): To determine the role of IP3R in HIV Gag release; (AIM 2): To determine the role of IP3R in RSV Gag release; (AIM 3): To test for IP3R-Gag interaction in cells and define the relationship to proteins known to be involved in endocytic trafficking. Genetic, biochemical, and cell biological approaches will be used that will include targeted protein depletion using microinjection and short hybrid interfering RNAs; targeted depletion coupled to replacement with mutated IP3R proteins; use of IP3R antagonists and agonists; and fluorescence-based approaches. This investigation will potentially link endocytic trafficking of retroviral Gag proteins to calcium signaling and provide new opportunities for development of novel antiviral strategies that interfere with virus assembly and transmission.

描述（由申请人提供）：1 型人类免疫缺陷病毒 (HIV-1) 是国内和国际大流行的病原体。目前还没有可用的疫苗，并且对当前基于药物的抗病毒疗法具有抵抗力的病毒变种正在世界范围内出现。这些事实使得确定抗病毒药物开发的新靶点和策略势在必行。 HIV和其他逆转录病毒的gag基因编码病毒结构前体多蛋白Gag，并且足以组装和释放病毒样颗粒。许多细胞蛋白有助于这一过程并代表潜在的新靶点。我们的初步研究表明，肌醇 (1,4,5)-三磷酸受体 (IP3R) 参与 HIV-1 Gag 和相关禽劳斯肉瘤病毒 (RSV) Gag 向质膜上释放位点的运输。此前尚未发现 IP3R 蛋白与逆转录病毒贩运有关。 IP3R 形成配体门控钙通道，调节细胞内钙储备的释放。钙释放调节许多细胞过程，包括细胞内运输和胞吐作用。 IP3R 抑制剂会干扰 Gag 贩运。对 IP3R 抑制剂的敏感性需要 HIV 和 RSV Gag 蛋白中的 Late 结构域，它们分别是结合 Tsg101 和 Nedd4 的区域。 Tsg101 和 Nedd4 是细胞蛋白，可分别促进 HIV 和 RSV Gag 运输至质膜上的位点，在质膜上 Gag 组装成病毒样颗粒 (VLP) 并出芽进入细胞外环境。拟议研究的目标是 (AIM 1)： 确定 IP3R 在 HIV Gag 释放中的作用； (目标 2)：确定 IP3R 在 RSV Gag 释放中的作用； (目标 3)：测试细胞中的 IP3R-Gag 相互作用，并确定与已知参与内吞运输的蛋白质的关系。将使用遗传、生物化学和细胞生物学方法，包括使用显微注射和短杂交干扰 RNA 进行靶向蛋白质消耗；靶向清除与突变 IP3R 蛋白替代相结合；使用IP3R拮抗剂和激动剂；和基于荧光的方法。这项研究可能会将逆转录病毒 Gag 蛋白的内吞运输与钙信号传导联系起来，并为开发干扰病毒组装和传播的新型抗病毒策略提供新的机会。