HIV FRAMESHIFTING--FROM BIOLOGY TO THERAPEUTICS

HIV框架转移——从生物学到治疗学

• 批准号: 2798211
• 负责人: STUART W PELTZ
• 金额: $ 12.44万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2000-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2798211
• 关键词: antiAIDS agent; antineoplastic antibiotics; computer assisted sequence analysis; drug resistance; drug screening /evaluation; frameshift mutation; gene induction /repression; genetic translation; human immunodeficiency virus 1; microorganism culture; nucleic acid sequence; ribosomes; virus genetics; virus replication

The ability of ribosomes to maintain the correct translational reading frame is fundamental to the integrity of protein synthesis and to cell growth and viability. However, there are now a number of examples utilized by viruses in which elongating ribosomes are programmed to shift their translational reading frame one base in the 5' direction. This process is called programmed -1 ribosomal frameshifting. Programmed -1 ribosomal frameshifting is utilized uniquely by eucaryotic viruses, making it a compelling target for developing antiviral agents. The human immunodeficiency virus type 1 (HIV-1) utilizes a programmed -1 ribosomal frameshift to synthesize both the gag and gag-pol proteins from a single transcript. We have been investigating the cis-acting elements and trans-acting factors that determine programmed -1 ribosomal frameshifting efficiencies in the yeast Saccharomyces cerevisiae. Using the double-stranded L-A virus system, we have shown that small changes in the ratio of the gag to gag-pol synthesized by altering frameshifting efficiencies leads to a loss of the killer virus. These findings have led us to develop the concept that antiviral agents can be identified that alter the efficiency of programmed frameshifting without dramatically affecting global protein synthesis. We have successfully demonstrated this principle using the yeast killer virus system as the model, and more recently, with HIV in mammalian cells. Based on these investigations, we propose to characterize programmed -1 frameshifting in HIV-1 with the goal of developing this virus specific mechanism as a target for antiviral intervention. The aims of the experiments proposed in this grant proposal will be to characterize the sequences that promote efficient frameshifting in HIV-1 and to determine the affects of altering frameshifting efficiency on HIV production. We will also determine if putative compounds that alter programmed -1 frameshifting and promote loss of the killer virus will also reduce or eliminate HIV production. Additional compounds that affect programmed frameshifting will also be identified. Finally, we will investigate at the molecular level how compounds that affect programmed frameshifting function. Our long term goal will be to develop compounds that affect frameshifting to the point that proof of principle has been established and antiviral agents targeting this process will be subsequently developed for clinical use.

核糖体保持正确的翻译阅读框的能力是蛋白质合成以及细胞生长和生存力的完整性的基础。 但是，现在有许多示例被病毒使用，其中伸长核糖体被编程为在5'方向上将其翻译阅读框架的一个基础移动。 此过程称为编程-1核糖体框架。 编程的-1核糖体框架是由桉树病毒独特地利用的，使其成为发展抗病毒药的引人注目的目标。 人类免疫缺陷病毒1型（HIV-1）利用一个编程的-1核糖体帧汇总来合成单个转录本的GAG和GAG-POL蛋白。 我们一直在研究确定酵母酵母酿酒酵母中编程的-1核糖体帧效率的顺式作用元件和跨作用因子。 使用双链L-A病毒系统，我们表明，通过改变Frameshiftifiesies的GAG与GAG-POL合成的GAG与POL的比率发生了较小的变化，从而导致杀伤病毒的丧失。 这些发现使我们发展了一个概念，即可以确定抗病毒剂可以改变编程帧的效率而不会显着影响全球蛋白质合成。 我们使用酵母菌病毒系统作为模型成功证明了这一原理，最近在哺乳动物细胞中使用HIV。 基于这些研究，我们建议在HIV -1中表征编程的-1框架，目的是开发该病毒特定机制作为抗病毒干预的靶标。 本赠款建议中提出的实验的目的是表征促进HIV-1中有效帧速率的序列，并确定改变弗拉米希效率对HIV产生的影响。我们还将确定假定的化合物是否改变了程序的-1帧速率并促进杀伤病毒的丧失也将减少或消除HIV产生。 还将确定影响编程帧的其他化合物。 最后，我们将在分子水平上研究如何影响编程帧函数的化合物。 我们的长期目标是开发影响框架的化合物，以至于已经建立了原理证明，并且针对该过程的抗病毒剂将随后开发用于临床使用。