Regulation of HIV-1 RNA Processing

HIV-1 RNA 加工的调控

• 批准号: 6989731
• 负责人: MASSIMO CAPUTI
• 金额: $ 32.51万
• 依托单位: FLORIDA ATLANTIC UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-15 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6989731
• 关键词: Regulation; HIV; RNA; Processing

DESCRIPTION (provided by applicant): The integrated HIV-1 proviral genome is transcribed by the host transcription machinery into a single 9.2 kb primary transcript. To express the nine different gene products required for viral replication, HIV has developed a number of strategies to regulate splicing and to circumvent cellular mechanisms restricting unspliced RNA in the cytoplasm. The regulatory protein Rev promotes transport of the incompletely spliced viral RNAs to the cytoplasm, binding to the Rev Responsive Element (RRE) on the viral RNA. Mechanisms regulating the appearance of unspliced and spliced viral RNAs, and their transport to the cytoplasm, involve interactions between viral RNA sequences and host cell factors. The characterization of these viral/host interactions could allow the design of novel therapeutic compounds. hnRNP H proteins have been shown to be necessary for HIV-1 RNA splicing in a series of in vitro experiments. They may also regulate RNA export and trafficking in infected cells. The role of single members of the hnRNP H protein family in HIV-1 RNA processing in vivo will be investigated by modulating hnRNP H protein expression in cells transfected with HIV-1-derived plasmids, bearing mutations in regulatory sequences. Several viral sequences that regulate viral RNA splicing in vitro have been identified. These will be tested to determine whether they are necessary for the regulation of viral RNA splicing in vivo. Preliminary data indicates that viral sequences interact with the RRE-Rev complex and, possibly, have a role in the Rev-dependent viral RNA transport pathway. These interactions will be characterized by a combination of in vivo and in vitro assays. Transcription and splicing have been shown to be closely coupled in other systems. Preliminary data indicates that the viral promoter influences splicing of viral substrates in vitro. Possible coupling of HIV-1 transcription and splicing will be investigated, using both a coupled transcription-splicing assay and transient transfection experiments.

描述（由申请人提供）：整合的 HIV-1 原病毒基因组由宿主转录机器转录成单个 9.2 kb 初级转录物。为了表达病毒复制所需的九种不同基因产物，HIV 开发了多种策略来调节剪接并规避限制细胞质中未剪接 RNA 的细胞机制。调节蛋白 Rev 促进不完全剪接的病毒 RNA 转运至细胞质，与病毒 RNA 上的 Rev 响应元件 (RRE) 结合。调节未剪接和剪接病毒 RNA 的出现及其向细胞质转运的机制涉及病毒 RNA 序列与宿主细胞因子之间的相互作用。这些病毒/宿主相互作用的特征可以帮助设计新型治疗化合物。在一系列体外实验中，hnRNP H 蛋白已被证明是 HIV-1 RNA 剪接所必需的。它们还可能调节受感染细胞中的 RNA 输出和运输。 hnRNP H 蛋白家族的单个成员在 HIV-1 RNA 体内加工中的作用将通过调节 hnRNP H 蛋白在用 HIV-1 衍生质粒转染的细胞中的表达来研究，该质粒在调节序列中带有突变。已经鉴定出几种在体外调节病毒RNA剪接的病毒序列。这些将被测试以确定它们对于体内病毒RNA剪接的调节是否是必需的。初步数据表明，病毒序列与 RRE-Rev 复合物相互作用，并且可能在 Rev 依赖性病毒 RNA 转运途径中发挥作用。这些相互作用将通过体内和体外测定的组合来表征。转录和剪接已被证明在其他系统中紧密耦合。初步数据表明，病毒启动子影响体外病毒底物的剪接。将使用偶联转录剪接测定和瞬时转染实验来研究 HIV-1 转录和剪接的可能偶联。