HIV PARTICLE FORMATION AND RNA PACKAGING

HIV 颗粒形成和 RNA 包装

• 批准号: 6374425
• 负责人: DAVID M REKOSH
• 金额: $ 25.56万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-15 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6374425
• 关键词: endoplasmic reticulum; gene expression; human immunodeficiency virus; intracellular transport; messenger RNA; ribosomes; virus RNA; virus assembly; virus protein

Description (Adapted from Applicant's abstract): The long-term goal of the research described in this proposal is to elucidate fundamental aspects of HIV virion formation and RNA packaging. Although the outline of the process of assembly has been known for some time, and closely follows that of the simpler retroviruses, many unanswered questions remain. Elucidating the mechanism of HIV viral particle formation and RNA packaging in more detail is likely to help identify key events that could become targets for the next generation of anti-HIV therapeutics. The applicant will address the following specific questions: Aim #1: Do assembly and RNA packaging interactions begin on the polyribosome? The applicant will explore this question by expressing Gagpol together with Gag, and independently from it on a different mRNA, to determine whether sythesis of Gagpol by frameshifting contributes to its incorporation into particles. The applicant will also address the issue of cis versus trans packaging of viral RNA. Aim #2: Do RNA localization and processing mechanisms contribute to HIV RNA packaging? Although specific packaging signals are known to exist within HIV genomic RNA, and the NC protien seems to play role in RNA recognition, little is known about other factors that facilitate packaging. In this aim the applicant will: a) determine if poly A plays a role in RNA packaging; b) determine if targeting an RNA to the surface of the endoplasmic reticulum affects its ability to be packaged; c) determine if the pathway of nuclear export affects packaging efficiency; d) determine if newly identified RNA trafficking signals (RTS) present in the HIV genome and "zipcodes" influence RNA packaging or assembly. Aim #3: What is the role of the 5' TAR element in RNA packaging and dimer linkage formation? When is the dimer linkage formed? The applicant has previously shown that the bottom part of the TAR stem is a necessary structural element for efficient RNA packaging. In this aim he will further explore this requirement, focusing attention on the possible role of TAR in dimer formation and dimer stability. He will also address the temporal order of dimer formation, i.e. can an RNA package without forming a dimer?

描述（改编自申请人的摘要）：该项目的长期目标 本提案中描述的研究旨在阐明艾滋病毒的基本方面 病毒体形成和RNA包装。虽然流程的概要 汇编已经为人所知有一段时间了，并且紧密遵循更简单的汇编 逆转录病毒，仍然有许多悬而未决的问题。阐明其机制 更详细地了解 HIV 病毒颗粒的形成和 RNA 包装可能会有所帮助 确定可能成为下一代目标的关键事件 抗艾滋病毒疗法。申请人将解决以下具体问题 问题： 目标#1：组装和 RNA 包装相互作用是否始于多核糖体？ 申请人将通过表达 Gagpol 以及 Gag，并独立于不同的 mRNA，以确定是否 通过移码合成 Gagpol 有助于将其纳入 颗粒。申请人还将解决顺式与反式的问题 病毒RNA的包装。 目标#2：RNA 定位和加工机制是否有助于 HIV RNA 包装？尽管已知 HIV 内存在特定的包装信号 基因组 RNA 和 NC 蛋白似乎在 RNA 识别中发挥作用，但很少 已知有利于包装的其他因素。为了这个目标 申请人将： a) 确定 Poly A 是否在 RNA 包装中发挥作用； b) 确定 RNA 是否靶向内质网表面 影响其包装能力； c) 确定核途径是否 出口影响包装效率； d) 确定是否有新鉴定的RNA HIV基因组中存在的贩运信号（RTS）和“邮政编码”的影响 RNA 包装或组装。 目标 #3：5' TAR 元件在 RNA 包装和二聚体中的作用是什么 连锁形成？二聚体连接何时形成？申请人有 先前表明，TAR 杆的底部是必要的结构 有效RNA包装的元件。为了这个目标，他将进一步探索这个 要求，重点关注 TAR 在二聚体形成中的可能作用 和二聚体稳定性。他还将讨论二聚体的时间顺序 形成，即RNA可以包装而不形成二聚体吗？