MECHANISMS OF ACTION OF INTERFERON

干扰素的作用机制

• 批准号: 3186792
• 负责人: MARIANO ESTEBAN
• 金额: $ 13.59万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-30 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3186792
• 关键词: adenosinetriphosphatase; antibody neutralization test; antiviral agents; cell free system; chemical fingerprinting; double stranded RNA; endoribonucleases; gel electrophoresis; gene expression; genetic manipulation; genetic markers; genetic transcription; genetic translation; immunogenetics; interferons; ligase; messenger RNA; molecular biology; molecular cloning; nucleic acid structure; oligonucleotides; phosphodiesterases; plasmids; protein kinase; thin layer chromatography; tissue /cell culture; vaccinia virus; virus DNA; virus replication

The antiviral and antiproliferative actions of interferons have been correlated with a number of enzyme activities (2-5A synthetase, endoribonuclease, protein kinase), which, when activiated, inhibit protein synthesis but the relevance of these enzymes to the inhibition of replication of RNA and DNA-containing viruses has not been elucidated. We have found in a vaccinia virus infected mouse L cell system that the interferon-mediated inhibition of viral protein synthesis in vivo correlates with activation of the 2-5A synthetase/endonuclease and degradation of viral RNAs. In contrast, in a number of mouse and human cells of different origins, vaccinia protein synthesis is not inhibited by interferon and a novel phenomenon has been discovered where vaccinia products block the 2-5A synthetase and protein kinase activities. In this proposal, experiments are described using vaccinia virus as a model system to elucidate: a) the in vivo role of the 2-5A synthetase/endonuclease system in the interferon-mediated inhibition of vaccinia virus protein synthesis; b) the mechanism by which a DNA virus such as vaccinia escapes blockade by the interferon system. To determine the in vivo role of the 2-5A synthetase/endonuclease on protein synthesis we will establish to what extent the integrity of viral and cellular RNAs is related to a block of translation, if specific degradation of certain RNAs occurs and if these events are the result of the formation of viral RNA during the course of infection. To define how vaccinia virus escapes inhibition by the interferon-mediated enzyme activities, we will characterize the nature and mode of action of vaccinia products with interfering properties present in cell-extracts and virions. To further define the vaccinia products with blocking effects on interferon action we will examine whether viral genes introduced into cells by DNA-mediated gene transfer can overcome specific interferon-mediated enzyme activities. By examining these cell-systems where the interferon response of the cells may be controlled by vaccinia gene(s) we may provide the means to define the mechanisms responsible for inhibition of replication of various viruses as well as the antiproliferative actions of interferons.

干扰素的抗病毒和抗增殖作用已被证实 与许多酶活性相关（2-5A 合成酶、 核糖核酸内切酶、蛋白激酶），当被激活时，会抑制蛋白质 合成，但这些酶与抑制的相关性 含RNA和DNA的病毒的复制尚未阐明。 我们 在痘苗病毒感染的小鼠 L 细胞系统中发现 干扰素介导的体内病毒蛋白合成抑制 与 2-5A 合成酶/核酸内切酶的激活相关 病毒RNA的降解。 相比之下，在许多小鼠和人类中 不同来源的细胞，牛痘蛋白的合成不受抑制 干扰素和一种新现象被发现 产品阻断 2-5A 合成酶和蛋白激酶活性。 在这个 建议，使用痘苗病毒作为模型系统描述实验 阐明：a) 2-5A 合成酶/核酸内切酶的体内作用 干扰素介导的痘苗病毒蛋白抑制系统 合成; b) DNA 病毒（如痘苗病毒）的逃逸机制 干扰素系统的封锁。 确定体内作用 2-5A 合成酶/核酸内切酶对蛋白质合成的影响我们将确定什么 病毒和细胞 RNA 的完整性与以下区域相关： 翻译，如果某些 RNA 发生特异性降解，并且如果这些 事件是病毒RNA在过程中形成的结果 感染。 确定痘苗病毒如何逃脱抑制 干扰素介导的酶活性，我们将表征其性质和 具有干扰特性的痘苗产品的作用方式 细胞提取物和病毒颗粒。 为了进一步定义痘苗产品 对干扰素作用的阻断作用我们将检查病毒基因是否 通过DNA介导的基因转移引入细胞可以克服特定的 干扰素介导的酶活性。 通过检查这些细胞系统 其中细胞的干扰素反应可能由牛痘控制 基因我们可以提供方法来定义负责的机制 抑制多种病毒的复制以及 干扰素的抗增殖作用。