GENETICS OF HERPESVIRUS - HOST CELL INTERACTIONS

疱疹病毒遗传学 - 宿主细胞相互作用

• 批准号: 3127772
• 负责人: MYRON LEVINE
• 金额: $ 35.91万
• 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/3127772
• 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; plasmids; protein biosynthesis; regulatory gene; ribonucleotide reductase; ribonucleotides; temperature sensitive mutant; tissue /cell culture; transcription factor; 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.

控制市场的监管现象分析 单纯疱疹病毒的协调有序合成 I型病毒（HSV-1）特异性蛋白在感染期间持续 成为这项研究的目标。 两项新举措是 提出了 第一个是对以往研究的扩展， 糖蛋白C（gC）基因表达的调节， 基因 晚期基因表达需要基因的反式激活 两个立即早期基因ICP 4和ICP 27的产物，并具有 对用于表达的病毒DNA的合成有严格的要求。 控制gC转录的顺式作用调节元件是 一个15 bp的序列，以TATA盒为中心。 的要求 可以通过融合上游的 早期tk基因5 ′-10 ′的转录调节序列 gC TATA元素 病毒DNA合成的要求 后期基因表达将是这些实验的主要焦点。 能够提供DNA的最小转录信号 gC基因的复制非依赖性表达将是 测定 假设ICP 8，HSV-1的主要DNA 结合蛋白，作为晚期基因的一般抑制子 将测试DNA复制之前的表达。 使用 携带tk-gC嵌合基因的病毒和温度 α 27基因的敏感突变等位基因，ICP 27的作用 在后期基因表达将被探测。 已经启动了关于表达调节的研究 HSV-1特异性核糖核苷酸大亚基的基因 还原酶（RR 1）。 RR 1基因表现出两者的特征 立即早期和早期基因调控，与所有 其他HSV-1基因，可能是唯一的反式激活的ICP 0，而不是 ICP 4在感染过程中 假设将被测试，在 通过alphaTIF实现感染立即早期表达 和八聚体/TAATGARATTC元件，而早期表达是 通过ICP 0和下游调节信号完成 RR 1启动子 顺式作用信号对于 将定义α TIF和ICP 0的反式激活。 交换 RR 1启动子和ICP 4之间的顺式作用信号 应答性TK启动子应鉴定应答性TK启动子的序列。 ICP 0和ICP 4诱导。 实验被提议为 确定α 0基因产物是否是唯一的病毒 在早期阶段调节RR 1表达的反式激活因子 感染 最后，RR 1启动子突变病毒将被 在小鼠中测试对神经毒力和 从潜伏期建立和重新激活。