NUCLEOSOMES AND HSV GENE REGULATION

核小体和 HSV 基因调控

• 批准号: 2897210
• 负责人: THOMAS C HOLLAND
• 金额: $ 3.82万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2897210
• 关键词: chromatin; gene expression; genetic promoter element; genetic regulation; herpes simplex virus 1; mutant; nucleosomes; protein structure function; tissue /cell culture; transcription factor; virus DNA; virus genetics; virus infection mechanism; virus protein

Nucleosomes are histone multimers around which approximately 146 base pairs of DNA are wrapped. They serve at least two functions in eukaryotic cells: compacting the DNA into chromatin and exerting a general repressive effect on gene expression. Considerable attention has been focused recently on the mechanisms by which cells alter chromatin structure to allow the cellular transcriptional machinery to gain access to genes and their promoters. Nucleosomes have been reported to form on the genomes of mammalian viruses, e.g., SV40. In the case of Herpes Simplex Virus Type 1 (HSV-1), published reports indicate that latent virus DNA is nucleosome associated. On the other hand, it has been reported that nucleosomes do not form on HSV-1 during the lytic replication cycle. However, our data strongly suggest that, in fact, nucleosomes do form on HSV-1 DNA during lytic replication. This has important implications for understanding the mechanisms of HSV-1 gene regulation. The goal of this proposal is to characterize the interactions of nucleosomes with the viral genome. Three specific aims are proposed. In the first aim, quantitative assays will be used to determine the fraction of HSV-1 DNA that is nucleosome associated. In the second aim, the organization of nucleosomes in the vicinity of the viral promoters will be determined. In the final aim, the effect of viral regulatory proteins on promoter chromatin structure will be determined. It is anticipated that this work will substantially enhance our knowledge of HSV-1 gene regulation, with important implications for our understanding of HSV-1 latency and pathogenesis, development of novel antiviral drugs, and possible use of HSV-1 as a vector for gene therapy.

核小体是组蛋白多聚体，周围有大约 146 个碱基 DNA 对被包裹。 它们至少有两个功能： 真核细胞：将DNA压缩成染色质并发挥作用 对基因表达的普遍抑制作用。引起了相当大的关注 最近关注细胞改变染色质的机制 允许细胞转录机器进入的结构 基因及其启动子。 据报道核小体可形成 哺乳动物病毒的基因组，例如 SV40。 疱疹的情况 单纯性病毒 1 型 (HSV-1)，已发表的报告表明，潜伏 病毒DNA与核小体相关。 另一方面，已经 据报道，HSV-1 在裂解过程中不会形成核小体 复制周期。 然而，我们的数据强烈表明，事实上， 在裂解复制过程中，HSV-1 DNA 上确实会形成核小体。 这有 对理解 HSV-1 基因机制的重要意义 规定。 该提案的目标是描述 核小体与病毒基因组的相互作用。 三个具体目标 被提议。 在第一个目标中，定量分析将用于 确定与核小体相关的 HSV-1 DNA 的分数。 在 第二个目标是在核小体附近组织核小体 将确定病毒启动子。 最终达到的效果是 启动子染色质结构上的病毒调节蛋白将 决定。 预计这项工作将大大提高 我们对 HSV-1 基因调控的了解，对 我们对 HSV-1 潜伏期和发病机制的了解、 新型抗病毒药物以及HSV-1作为基因载体的可能用途 治疗。