GENETICS OF HERPESVIRUS - HOST CELL INTERACTIONS

疱疹病毒遗传学 - 宿主细胞相互作用

• 批准号: 3127771
• 负责人: MYRON LEVINE
• 金额: $ 34.59万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3127771
• 关键词: DNA binding protein; DNA replication; Herpes simplex disease; RNA; chromosome deletion; electrophoresis; gene expression; genetic promoter element; genetic transcription; glycoproteins; herpes simplex virus 1; laboratory mouse; latent virus infection; nucleic acid hybridization; nucleic acid sequence; oxidoreductase; plasmids; protein biosynthesis; regulatory gene; ribonucleotides; temperature sensitive mutant; tissue /cell culture; transfection; virus genetics; virus infection mechanism; virus replication

The analysis of the regulatory phenomena controlling the coordinately and sequentially ordered synthesis of herpes simplex virus type I (HSV-1) specific proteins during infection continues to be the objective of this research. Two new initiatives are proposed. The first is an extension of previous studies on the regulation of expression of the glycoprotein C (gC) gene, a late gene. Late gene expression requires transactivation by the gene products of two immediate-early genes, ICP4 and ICP27, and has a strict requirement for the synthesis of viral DNA for expression. The cis-acting regulatory element controlling gC transcription is a 15 bp sequence centered around the TATA box. The requirement for DNA synthesis can be alleviated by fusing the upstream transcriptional regulatory sequences of the early tk gene 5' to the gC TATA element. The requirement for viral DNA synthesis for late gene expression will be the main focus of these experiments. The minimum transcriptional signal capable of giving DNA replication independent expression of the gC gene will be determined. The hypothesis that ICP8, the HSV-1 major DNA binding protein, functions as a general repressor of late gene expression prior to DNA replication will be tested. Using a virus carrying the tk-gC chimeric gene and a temperature sensitive mutant allele of the alpha 27 gene, the role of ICP27 in late gene expression will be probed. Studies have been initiated on the regulation of expression of the gene for the HSV-1 specified large subunit of ribonucleotide reductase (RR1). The RR1 gene shows characteristics of both immediate-early and early gene regulation and, in contrast to all other HSV-1 genes, may be uniquely transactivated by ICP0 instead of ICP4 during infection. The hypothesis will be tested that in infection immediate-early expression is achieved through alphaTIF and the octamer/TAATGARATTC element, while early expression is accomplished through ICP0 and the downstream regulatory signals of the RR1 promoter. The cis-acting signals critical for transactivation by alpha TIF and ICP0 will be defined. Exchange of cis-acting signals between the RR1 promoter and the ICP4 responsive tk promoter should identify the sequences responsive to induction by ICP0 and ICP4. Experiments are proposed to determine if the alpha0 gene product is the only viral transactivator regulating RR1 expression during the early phase of infection. Finally, the RR1 promoter mutant viruses will be tested in mice for effects on neurovirulence and on the establishment and reactivation from latency.

对我国证券市场管制现象的分析 单纯疱疹病毒的配位有序合成 感染期间I型单纯疱疹病毒(HSV-1)特异性蛋白继续存在 作为本研究的目标。两项新举措是 建议。第一种是对前人研究的延伸 晚期糖蛋白C(GC)基因的表达调控 吉恩。晚期基因表达需要基因反式激活 两个直接早期基因ICP4和ICP27的产物，具有 对合成表达的病毒DNA要求严格。 控制GC转录的顺式作用调控元件是 以塔塔框为中心的15个碱基的序列。这一要求 因为DNA的合成可以通过融合上游的 早期tk基因5‘T0转录调控序列 GC塔塔元素。对病毒DNA合成的要求 晚期基因表达将是这些实验的主要焦点。 能够给出DNA的最低转录信号 GC基因的复制无关表达将是 下定决心。ICP8，HSV-1主要DNA的假说 结合蛋白--晚期基因的一般抑制因子 将对DNA复制前的表达进行测试。使用 携带tk-gc嵌合基因和温度的病毒 α27基因的敏感突变等位基因ICP27的作用 后期将对基因表达进行探讨。 已有研究开始研究对基因表达的调控 单纯疱疹病毒1型特异性核糖核酸大亚基基因 还原酶(RR1)。RR1基因表现出两者的特征 即刻-早期和早期的基因调控，与所有 其他HSV-1基因可能被ICP0唯一反式激活 ICP4在感染过程中的表达。这一假设将得到检验，在 感染的即刻-早期表达通过alphaTIF实现 和八聚体/TAATGARATTC元件，而早期表达是 通过ICP0和下游调节信号实现 RR1启动子的序列。顺式作用信号对 将定义阿尔法TIF和ICP0的激活。交易所 RR1启动子和ICP4之间的顺式作用信号 有反应的tk启动子应该识别有反应的序列 对ICP0和ICP4的诱导作用。建议进行实验，以 确定alpha0基因产物是否是唯一的病毒 反式激活剂在早期调节RR1的表达 感染的可能性。最后，RR1启动子突变病毒将被 在小鼠身上测试对神经毒力和对 从延迟中建立和重新激活。