HIV Genomic RNA Structure

HIV 基因组 RNA 结构

• 批准号: 7188632
• 负责人: J. Stephen Lodmell
• 金额: $ 22.9万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-15 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7188632
• 关键词: Adopted; Affect; Anti-Retroviral Agents; Antibiotics; Base Pairing; Behavior; Binding; Biochemical; Biochemical Genetics; Biochemistry; Biological Assay; Cellular biology; Code; Cultured Cells; Deletion Mutation; Dimerization; Disruption; Drug Delivery Systems; Elements; Event; Functional RNA; Gagging; Genes; Genetic Transcription; Genomics; HIV; HIV-1; HIV-2; Human; In Vitro; Isomerism; Molecular Conformation; Mutation; Personal Satisfaction; Phylogenetic Analysis; Process; Protein Biosynthesis; RNA; RNA Sequences; RNA Splicing; Range; Regulation; Research Personnel; Retroviridae; Ribosomal RNA; Role; Signal Transduction; Site; Staging; Structure; Subfamily lentivirinae; Testing; Transactivation; Translation Initiation; Translations; Viral; Viral Proteins; Virus; base; design; gag Gene Products; mutant; prevent; programs; stem; viral RNA

DESCRIPTION (provided by applicant): The 5' untranslated leader region of HIV-1 and HIV-2 genomic RNA contains sequences essential for viral replication, including signals for transactivation of transcription, primer binding, splicing, encapsidation, RNA dimerization, and initiation of translation of the gag gene. Within a species, phylogenetic conservation of these non-coding sequences rivals even that of conserved coding domains for the viral proteins, attesting to their importance for replication. The high degree of conservation also highlights the potential to exploit this region for antiretroviral strategies. The use of conserved RNA sequences as drug targets has been well established, since many clinically useful antibiotics recognize a specific segment of bacterial rRNA. Biochemical characterization of the leader RNA of HIV-1 and HIV-2 suggests that it adopts multiple conformations that alternately display or hide RNA signals appropriate to different stages of the replication cycle. Based on the in vitro characterization of conformational isomers of HIV-2 leader RNA and upon the high degree of conservation of involved sequences, Dr. Lodmell hypothesizes that the leader region of HIV and other retroviruses is intimately involved in the regulation of several viral replicative processes, including dimerization of viral RNA, encapsidation, splicing, and protein synthesis, and that this regulation is manifested by differential presentation of the signaling structures found in the leader region. In this application, Dr. Lodmells proposes 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. A combination of cell culture, genetic, and biochemical approaches will be used to characterize the largely unexplored interrelationships of RNA splicing, dimerization, encapsidation, and translation in HIV types 1 and 2. The studies proposed here will aid in our overall understanding of the biochemistry and cell biology of human lentivirus replication as well as to validate the 5' leader region RNA as a potential antiretroviral target.

描述（由申请人提供）：HIV-1和HIV-2基因组RNA的5'非翻译前导区含有病毒复制所必需的序列，包括转录反式激活、引物结合、剪接、衣壳化、RNA二聚化和gag基因翻译起始的信号。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的保守编码域相媲美，证明了它们对于复制的重要性。高度的保护也凸显了利用该地区进行抗逆转录病毒策略的潜力。使用保守的 RNA 序列作为药物靶标已经很成熟，因为许多临床上有用的抗生素可以识别细菌 rRNA 的特定片段。 HIV-1和HIV-2前导RNA的生化特征表明，它采用多种构象，交替显示或隐藏适合复制周期不同阶段的RNA信号。基于 HIV-2 前导 RNA 构象异构体的体外表征以及相关序列的高度保守性，Lodmell 博士假设 HIV 和其他逆转录病毒的前导区域密切参与多种病毒复制过程的调节，包括病毒 RNA 的二聚化、衣壳化、剪接和蛋白质合成，并且这种调节通过前导区的差异表达来体现。 在前导区发现信号结构。在本申请中，Lodmells 博士建议通过构建和表征含有替换和缺失突变的病毒突变体来测试这一假设，这些突变体旨在干扰长程和短程 RNA 相互作用，这些相互作用在体外被证明对 RNA 结构和行为的调节很重要。细胞培养、遗传和生化方法的结合将用于表征 1 型和 2 型 HIV 中 RNA 剪接、二聚化、衣壳化和翻译的很大程度上尚未探索的相互关系。此处提出的研究将有助于我们全面了解人类慢病毒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药物 目标。