Regulation of HIV-1 RNA Processing

HIV-1 RNA 加工的调控

• 批准号: 6688992
• 负责人: MASSIMO CAPUTI
• 金额: $ 24.59万
• 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/6688992
• 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）：综合HIV-1病毒基因组由主机转录机械转录为单个9.2 kb主转录本。为了表达病毒复制所需的九种不同的基因产品，HIV制定了许多策略来调节剪接和绕过细胞限制细胞质中未植物RNA的细胞机制。调节蛋白REV促进了不完全剪接的病毒RNA向细胞质的转运，并与病毒RNA上的Rev Responsive元件（RRE）结合。调节未贴合和剪接病毒RNA的出现的机制及其转运到细胞质，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/宿主相互作用的表征可以允许设计新颖的治疗化合物。 HNRNP H蛋白已被证明是在一系列体外实验中进行HIV-1 RNA剪接所必需的。它们还可能调节RNA的导出和运输感染细胞。 HNRNP H蛋白家族中单个成员在HIV-1 RNA加工体内的作用将通过调节用HIV-1衍生的质粒转染的细胞中调节HNRNP H蛋白表达，并在调节序列中携带突变。已经鉴定出几种调节病毒RNA剪接的病毒序列。这些将进行测试以确定它们是否对于在体内调节病毒RNA剪接是否必要。初步数据表明，病毒序列与RRE-REV复合物相互作用，并且可能在依赖Rev的病毒RNA转运途径中起作用。这些相互作用的特征是体内和体外测定的结合。转录和剪接已显示在其他系统中紧密耦合。初步数据表明，病毒启动子在体外影响病毒底物的剪接。将研究HIV-1转录和剪接的可能耦合，使用耦合的转录分解测定和瞬态转染实验。