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架的能力是蛋白质合成的完整性以及细胞生长和存活的基础。然而，现在有许多病毒利用的例子，在这些例子中，延长的核糖体被编程为将其翻译阅读框架向5‘方向移动一个碱基。这一过程被称为程序性核糖体移码。程序性核糖体移码被真核病毒所独有，使其成为开发抗病毒药物的一个引人注目的靶点。人类免疫缺陷病毒1型(HIV-1)利用程序化的-1核糖体移码从单个转录本合成Gag和Gag-Poll蛋白。我们一直在研究决定酿酒酵母中程序性核糖体移码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，我们证明了通过改变移帧效率而合成的Gag/Gag-Poll的比例的微小变化会导致杀手病毒的损失。这些发现使我们形成了这样的概念，即可以确定抗病毒药物可以改变程序移码的效率，而不会显著影响全球蛋白质合成。我们已经成功地用酵母杀手病毒系统作为模型，并在最近用哺乳动物细胞中的艾滋病毒证明了这一原理。在这些研究的基础上，我们建议对HIV-1中的程序性-1移码进行表征，目的是开发这种病毒特异性机制作为抗病毒干预的目标。这项拨款提案中提出的实验的目的将是描述促进HIV-1中有效移码的序列的特征，并确定改变移码效率对HIV产生的影响。我们还将确定改变程序化移码和促进杀手病毒丢失的假定化合物是否也会减少或消除HIV的产生。还将确定影响程序性移码的其他化合物。最后，我们将在分子水平上研究影响程序性移码的化合物是如何发挥作用的。我们的长期目标将是开发影响移码的化合物，以确保已经建立了原理证据，并将随后开发针对这一过程的抗病毒药物用于临床。