ROLE OF BKV ENHANCERS IN VIRUS REGULATION AND CANCER

BKV 增强剂在病毒调节和癌症中的作用

• 批准号: 3187861
• 负责人: ROBERT Paul RICCIARDI
• 金额: $ 12.61万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3187861
• 关键词: DNA binding protein; Polyomavirus; chromosome inversion; gene deletion mutation; gene duplication; genetic enhancer element; genetic manipulation; genetic promoter element; genetic transcription; hamsters; lysogeny; molecular cloning; nucleoproteins; plasmids; point mutation; viral carcinogenesis; virus DNA; virus genetics; virus replication

Our aim is to define the details of how the enhancers of the human virus, BKV, regulate the biology of the virus and to determine what role the BKV enhancers play in viral transformation and tumorigenesis. The enhancer element of BKV consists of three 68 bp tandem repeats which stimulate early viral promoter transcription. Recently, we have discovered that the BKV enhancers are necessary for viral replication. Also, reducing the number of BKV enhancers is known to increase the transformation efficiency of the virus. By extrapolation, it is likely that the BKV enhancers affect episomal persistence of the viral genome and tumorigenesis. Our approach will be to generate a variety of different enhancer mutations and assay for altered viral regulation, transformation and tumorigenesis. The types of enhancer element mutations that will be generated are: enhancer elements with one, and two repeats; 5' and 3' enhancer terminal deletions; internal deletions; site-specific mutations of interesting landmark sequences; reverse orientation; and direct substitution of BKV enhancers with those of other viruses. We will test the enhancer mutations both in the context of virus and plasmids, in which specific viral functions can be studied independently of one another. These viruses and plasmids will be tested in distinct assay systems to determine the effect of the mutated enhancers on viral replication, transcription, episomal persistence, transformation and tumorigenesis. We will also use a variety of techniques to explore the binding of specific proteins to the BKV enhancers with the aim of defining how these proteins may regulate the different enhancer functions. In particular, we will investigate the role of the TGGCA nuclear protein which specifically binds to the BKV enhancers. We will use our site- specific enhancer mutants to define the relationship between altered replication and binding of the TGGCA protein. We will also attempt to develop an in vitro replication system to further define the role of the BKV enhancers in replication.

我们的目的是定义有关如何增强器的细节 人类病毒，BKV调节病毒的生物学 确定BKV增强剂在病毒中的作用 转化和肿瘤发生。 BKV的增强子元素 由三个68 bp串联重复组成，这些重复刺激早期病毒 启动子转录。 最近，我们发现 BKV增强子是病毒复制所必需的。 另外，还原 已知BKV增强剂的数量增加了 病毒的转化效率。 通过外推，是 BKV增强子可能会影响 病毒基因组和肿瘤发生。 我们的方法是生成 各种不同的增强子突变和更改的测定 病毒调节，转化和肿瘤发生。 类型 将生成的增强子元素突变是：增强剂 带有一个和两个重复的元素； 5'和3'增强器终端 删除；内部删除；有趣的现场突变 地标序列；反向方向；和直接替代 带有其他病毒的BKV增强子的图。 我们将测试 在病毒和质粒的背景下，增强子突变，在 可以独立研究哪些特定病毒功能 其他。 这些病毒和质粒将在不同的 测定系统以确定突变增强器的效果 关于病毒复制，转录，偶发性持久性， 转化和肿瘤发生。 我们还将使用各种 探索特定蛋白与BKV的结合的技术 增强剂的目的是定义这些蛋白质如何 调节不同的增强子功能。 特别是，我们会 研究TGGCA核蛋白的作用，该蛋白质的作用 特别结合BKV增强器。 我们将使用我们的网站 - 特定增强子突变体以定义 TGGCA蛋白的复制和结合改变。 我们将 还尝试开发体外复制系统以进一步 定义BKV增强子在复制中的作用。