HIV-2 Genomic RNA Structure

HIV-2 基因组 RNA 结构

• 批准号: 6843034
• 负责人: J. Stephen Lodmell
• 金额: $ 23.97万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2005-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6843034
• 关键词: RNA; RNA splicing; capsid; gene deletion mutation; gene mutation; genetic translation; human immunodeficiency virus 2; nucleic acid repetitive sequence; nucleic acid structure; recombinant virus; tissue /cell culture; virus genetics; virus replication

DESCRIPTION (provided by applicant): The 5' leader region of retroviral genomic RNA, while not translated, is replete with other non-coding functionality, including sequences important for transactivation of transcription, primer binding, splicing, encapsidation, dimerization of viral RNA, and initiation of translation of the gag ORF. Within a species, phylogenetic conservation of these non-coding sequences rivals even that of conserved coding domains for the viral proteins. This observation emphasizes that the sequence and structure of the leader region are essential for viral replication. The high degree of conservation also highlights the potential to target this region for antiretroviral strategies. The use of conserved RNA sequences as drug targets have been well established, since many ribosome-targeting antibiotics recognize a specific segment of bacterial rRNA. Recent in vitro results have demonstrated that structure of the leader RNA is plastic by design and its multiple conformations appear to give rise to functions appropriate to the stage of the replication cycle. In the last grant period, we elucidated structural and dynamic aspects of the leader region of the genomic RNA of HIV-2, one of the causative agents of AIDS in humans. In particular, while searching for signals important for genomic RNA dimerization, we discovered that these signals were interspersed with and overlaped with known signals for encapsidation, splicing, and translation of viral RNAs. Based on the in vitro characterization of conformational isomers of HIV-2 leader RNA and upon the high degree of phylogenetic conservation of involved sequences, we hypothesize that the leader region of HIV-2 and other retroviruses is intimately involved in the regulation of several viral processes, and that this regulation is manifested by differential presentation of the signaling structures found in the leader region. In this application we propose to test this hypothesis by constructing and characterizing viral mutants harboring substitution and deletion mutations designed to interfere with long- and short-range RNA interactions that were shown in vitro to be important in the modulation of RNA structure and behavior. We will use a combination cell culture and biochemical approaches to characterize the largely unexplored interrelationships of RNA dimerization, encapsidation, and translation in HIV-2. The studies proposed here will aid in our overall understanding of the biochemistry and cell biology of HIV-2 replication as well as to validate the 5' leader region RNA as a potential antiretroviral target.

描述（由申请人提供）：逆转录病毒基因组RNA的5'前导区虽然不翻译，但充满了其他非编码功能，包括对转录的反式激活、引物结合、剪接、腺苷酸化、病毒RNA的二聚化和gag ORF翻译的起始重要的序列。在一个物种内，这些非编码序列的系统发育保守性甚至可以与病毒蛋白质的保守编码结构域相媲美。这一观察结果强调了前导区的序列和结构对于病毒复制是必不可少的。高度的保护也突出了针对这一地区的抗逆转录病毒战略的潜力。由于许多核糖体靶向抗生素识别细菌rRNA的特定片段，因此保守RNA序列作为药物靶标的使用已经得到很好的建立。 最近的体外研究结果表明，前导RNA的结构是塑料的设计和它的多种构象出现，产生适当的复制周期的阶段的功能。在上一个资助期间，我们阐明了HIV-2基因组RNA前导区的结构和动力学方面，HIV-2是人类艾滋病的病原体之一。特别是，在寻找对基因组RNA二聚化重要的信号时，我们发现这些信号与病毒RNA的剪接、剪接和翻译的已知信号穿插在一起。基于体外表征的构象异构体的HIV-2的前导RNA和高度的系统发育保守的相关序列，我们假设，HIV-2和其他逆转录病毒的前导区密切参与的几个病毒过程的调节，这种调节表现为差异介绍的前导区中发现的信号结构。 在本申请中，我们建议通过构建和表征病毒突变体来测试这一假设，所述病毒突变体具有设计用于干扰长距离和短距离RNA相互作用的取代和缺失突变，所述长距离和短距离RNA相互作用在体外显示出在RNA结构和行为的调节中是重要的。我们将使用细胞培养和生物化学方法相结合的特点，在很大程度上未开发的相互关系的RNA二聚化，双糖苷化，和翻译的HIV-2。本文提出的研究将有助于我们全面了解HIV-2复制的生物化学和细胞生物学，并验证5'前导区RNA作为潜在的抗逆转录病毒靶点。