Molecular Interactions Between Rev and Cellular Factors

Rev 和细胞因子之间的分子相互作用

• 批准号: 7759611
• 负责人: Maria L. Zapp
• 金额: $ 30.65万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-15 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7759611
• 关键词: Ablation; Antiviral Agents; Binding; Biochemical Genetics; Biological Assay; Cell Line; Cell Nucleus; Cell Survival; Cells; Characteristics; Complex; Cytoplasm; Development; Energy Transfer; Event; Fluorescence; Funding; Gagging; Goals; HIV; HIV Budding; HIV-1; Human; Introns; Life; Life Cycle Stages; Link; Mammalian Cell; Mediating; Methods; Molecular; Movement; Nuclear Export; Nuclear RNA; Pathway interactions; Production; Proteins; RNA; RNA Interference; Reproduction; Role; Specificity; Viral; Viral Regulatory Proteins; Virus; Virus Assembly; base; cofactor; drug development; human HRB protein; macrophage; protein expression; research study; rev Protein; tissue/cell culture; trafficking; viral RNA

Replication of human immunodeficiency virus type 1 (HIV-1) entails a tightly regulated, complex life cycle, which is dependent upon specific interactions between viral RNAs and viral and cellular proteins. An essential and characteristic step in the viral replication cycle is the export of the intron-containing gag-pol and env mRNAs from the nucleus to the cytoplasm. The viral regulatory protein Rev mediates this event, in conjunction with the cellular nuclear export machinery and several protein cofactors. Cellular factors that are essential for Rev-directed RNA export offer alternate targets for inactivation and are potential candidates for antiviral drug development. During the past funding period, we have implicated the human Rev Interacting Protein (hRIP) and other cellular factors in a protein interaction network by which Rev mediates export of HIV RNAs from the nucleus to the cytoplasm. Our studies have revealed that hRIP is an essential cellular HIV cofactor that acts at a previously unanticipated step in Rev-directed RNA export: release of RNAs from the nuclear periphery to the cytoplasm. This perinuclear step may link nuclear export of Rev-directed RNAs with their cytoplasmic localization and function, in particular, virus assembly. Experiments are proposed to delineate the physical and functional protein interaction network mediating Rev-directed RNA export in living cells. We will use biochemical, genetic, and cell-based approaches to continue to study the function of hRIP and analyze how hRIP promotes movement of HIV RNAs from the nuclear periphery. Additionally, our studies of hRIP led us to investigate interactions between Rev and proteins that mediate cellular trafficking pathways. Experiments are proposed to explore the possibility of a functional link between Rev-directed RNA export and cellular trafficking pathways involved in HIV budding and assembly. hRIP is not required for cell viability and thus, may be an attractive target for the development of new antiviral strategies. During the past funding period, we have shown that ablation of hRIP activity using RNA interference (RNAi) results in the loss of HIV replication in mammalian cells. We will use several approaches to optimize the efficiency and specificity of RNAi-mediated silencing of hRIP expression and validate its inhibitory effect on HIV replication in mammalian cell lines and primary cells. The ultimate goal of these experiments is to validate cellular hRIP activity as a target for new antiviral strategies.

人类免疫缺陷病毒1型（HIV-1）的复制需要严格调节的复杂生命周期， 这取决于病毒RNA与病毒和细胞蛋白之间的特定相互作用。一个 病毒复制周期中的基本和特征步骤是导出含含有器的GAG-POL 以及从细胞核到细胞质的mRNA。病毒调节蛋白Rev介导了此事件， 与细胞核输出机械和几种蛋白质辅因子的结合。细胞因素是 对于转速导向的RNA出口至关重要，提供替代目标，以使其失活，并且是潜在的候选者 抗病毒药物开发。在过去的资金期间，我们牵涉到人类的Rev互动 蛋白质（HRIP）和其他细胞因子在蛋白质相互作用网络中介导的出口 HIV RNA从细胞核到细胞质。我们的研究表明，HRIP是必不可少的细胞 艾滋病毒辅助因子在以前意外的RED指导RNA导出的步骤中起作用：从RNA中释放 细胞质的核外围。这个核周步骤可能会连接转化为RNA的核出口 其细胞质定位和功能，尤其是病毒组装。提出了实验 描述介导Rev-direction的RNA导出的物理和功能性蛋白质相互作用网络 细胞。我们将使用生化，遗传和基于细胞的方法继续研究HRIP的功能 并分析HRIP如何促进HIV RNA从核外围的运动。另外，我们的 HRIP的研究使我们研究了介导细胞运输的Rev和蛋白质之间的相互作用 途径。提出了实验以探索Rev-Direction的功能联系的可能性 RNA出口和细胞运输途径涉及HIV萌芽和组装。不需要HRIP 细胞活力，因此，可能是发展新抗病毒策略的有吸引力的目标。在 过去的资金期，我们已经表明，使用RNA干扰（RNAI）消融HRIP活性导致 哺乳动物细胞中HIV复制的丧失。我们将使用几种方法来优化效率 RNAi介导的HRIP表达沉默的特异性并验证其对HIV的抑制作用 在哺乳动物细胞系和原代细胞中复制。这些实验的最终目标是验证 细胞HRIP活性是新的抗病毒策略的目标。

项目成果

Consensus nomenclature for the human ArfGAP domain-containing proteins.

• DOI: 10.1083/jcb.200806041
• 发表时间: 2008-09-22
• 期刊: The Journal of cell biology
• 作者: Kahn RA;Bruford E;Inoue H;Logsdon JM Jr;Nie Z;Premont RT;Randazzo PA;Satake M;Theibert AB;Zapp ML;Cassel D
• 通讯作者: Cassel D