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.

核糖体维持正确的翻译阅读框的能力对蛋白质合成的完整性和细胞生长和活力至关重要。然而，现在有许多病毒使用的例子，其中延长核糖体被编程使其翻译阅读框在5'方向上移动一个碱基。这个过程被称为程序化-1核糖体移框。程序-1核糖体移框被真核病毒独特地利用，使其成为开发抗病毒药物的一个引人注目的目标。人类免疫缺陷病毒1型（HIV-1）利用程序化-1核糖体移码从单个转录物合成gag和gag-pol蛋白。我们一直在研究决定酿酒酵母程序-1核糖体移框效率的顺式作用元件和反式作用因子。利用双链L-A病毒系统，我们已经证明，通过改变移帧效率合成的gag与gag-pol的比例的微小变化会导致杀伤病毒的损失。这些发现使我们发展出一种概念，即抗病毒药物可以在不显著影响全局蛋白质合成的情况下改变程序化移框的效率。我们已经用酵母杀伤病毒系统作为模型成功地证明了这一原理，最近又用哺乳动物细胞中的HIV证明了这一原理。基于这些研究，我们建议描述HIV-1中的编程-1移帧，目的是开发这种病毒特异性机制作为抗病毒干预的靶点。本拨款提案中提出的实验目的将是表征促进HIV-1有效移帧的序列，并确定改变移帧效率对HIV产生的影响。我们还将确定，如果假定的化合物改变编程-1帧移位和促进杀伤病毒的损失，也会减少或消除艾滋病毒的产生。还将确定影响程序移帧的其他化合物。最后，我们将在分子水平上研究影响程序移框功能的化合物。我们的长期目标将是开发影响帧移的化合物，直至原理证明已经建立，针对这一过程的抗病毒药物将随后开发用于临床。