GENE EXPRESSION DURING HSV-1 LATENCY AND REACTIVATION

HSV-1 潜伏期和重新激活期间的基因表达

• 批准号: 6273863
• 负责人: NIGEL William FRASER
• 金额: $ 20.27万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6273863
• 关键词: Herpes simplex disease; PC12 cells; RNA splicing; disease /disorder model; gene deletion mutation; gene expression; genetic promoter element; genetically modified animals; genome; herpes simplex virus 1; human tissue; in situ hybridization; laboratory mouse; latent virus infection; neurotropic virus; phenotype; polymerase chain reaction; transcription factor; trigeminal nerve; virus DNA; virus RNA; virus genetics; virus protein; virus replication

Human neurotropic herpes viruses such as HSV cause much disease and suffering. The long term objective of this project is to understand the mechanism of herpes virus latency and reactivation at the molecular level. In the present application, we propose to continue to examine the latency associated transcripts (LATs) to determine what role they play in viral latency and reactivation. Their regulation through the cellular early growth response (EGR) proteins will also be studied. These goals will be achieved using the techniques of molecular virology. We will study expression of a newly identified latency associated transcription unit, which appears to map upstream of the well recognized LAT. This transcription unit is encoded in a region of the viral genome to which the slow or inefficient reactivation phenotype of some HSV mutants has been mapped, and we are presently defining this genetic locus. A second phenotype has recently been associated with the 2 kb LAT RNA. Mutants that are deleted in part of this transcript appear to form latency in a reduced number of neuronal cells. Key to the understanding of this phenotype will be knowledge of the mechanism underlying the cellular localization of the 2 kb LAT RNA. Our funding that the 2 kb LAT RNA is present in the cytoplasm during lytic infection and appears associated with ribosomal subunits suggests a role in translation for this RNA. In order to understand why the LAT RNA is present in the nucleus during latency and in the cytoplasm during reactivation and the lytic cycle of infection, we will examine the structure of the 2 kb LAT through comparisons of, the 5' end position, the presence of a cap, splicing, and promoter studies, between nuclear and cytoplasmic LAT species. In order to further study the 2 kb LAT and the role that its cellular distribution plays in the apparent reduction of latently infected cells, we will develop cell lines expressing 2 kb LAT. This will be done in fibroblasts, CV-1 cells, cells of neuronal origin, Pc12 cells and ES cells. Finally, from cell lines expressing 2 kb LAT in ES cells, transgenic animals will be made to study LAT expression and cellular localization after reactivation stimuli are applied. We will study the role of EGR proteins in modulating the LAT gene expression as once we understand what the LAT gene does, it will be important to understand how the LAT gene function is regulated.

人类神经疱疹病毒（例如HSV）引起了很多疾病， 痛苦。该项目的长期目标是了解 疱疹病毒潜伏期的机理和分子水平的重新激活。 在本应用程序中，我们建议继续检查潜伏期 相关的成绩单（LATS）来确定它们在病毒中扮演什么角色 潜伏期和重新激活。它们通过细胞早期调节 还将研究生长反应（EGR）蛋白。这些目标将是 使用分子病毒学技术实现。 我们将研究与新确定的延迟相关的表达 转录单元，似乎在公认的上游绘制 拉特。该转录单元编码在病毒基因组的区域中 某些HSV突变体的缓慢或效率低下的重新激活表型 已被映射，我们目前正在定义这个遗传基因座。 第二种表型最近与2 kb LAT RNA相关。 该成绩单中部分删除的突变体似乎形成了潜伏期 在减少的神经元细胞中。理解这一点的关键 表型将了解细胞的基础机制 2 kb lat RNA的定位。我们的资助是2 kb lat rna是 裂解感染期间的细胞质中存在，并且似乎相关 带有核糖体亚基表明在该RNA的翻译中起作用。 为了理解为什么在核中存在LAT RNA在 在重新激活过程中的潜伏期和细胞质中 感染，我们将检查2 kb lat的结构 比较5'端位置，盖子的存在，剪接和 核和细胞质LAT物种之间的启动子研究。 为了进一步研究2 kb的lat及其细胞的作用 分布在明显减少了潜在感染的细胞中， 我们将开发表达2 kb LAT的细胞系。这将在 成纤维细胞，CV-1细胞，神经元起源的细胞，PC12细胞和ES 细胞。最后，从表达ES细胞中2 kb LAT的细胞系， 将使转基因动物研究LAT表达和细胞 应用重新激活刺激后的定位。 我们将研究EGR蛋白在调节LAT基因中的作用 表达一旦我们了解LAT基因的作用，它将是 了解如何调节LAT基因功能。