Integration and Expression of Retrovirus DNA

逆转录病毒DNA的整合和表达

• 批准号: 6522715
• 负责人: JOHN M COFFIN
• 金额: $ 39.63万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6522715
• 关键词: Retroviridae disease; avian leukosis virus; enzyme activity; host organism interaction; integrase; metallothionein; mitochondrial DNA; provirus; tissue /cell culture; virus DNA; virus genetics; virus infection mechanism; virus integration

Integration of viral into host cell DNA is a central and unique feature of retrovirus replication, and is directly responsible for many of the special characteristics of this important group of viruses, including their ability to cause cancer by activating and transducing oncogenes, their evolutionary persistence as endogenous viruses in the host germline, their very broad pathogenic spectrum, and their ability to serve as vectors for the gene therapist. Although the biochemical and structural aspects of the integration process are becoming quite well worked out, many important features remain unknown, including what defines an integration target, the relationship between chromatin and DNA structure and integration specificity, and the effect of the location of the integrated provirus on its subsequent expression. The overall goal of this project is to improve our understanding of key aspects of the role of integration in the virus-host interaction. Specifically, we will address the following questions: 1. Can we define an optimal target sequence (or structure) for integrase in vitro? What are the structural correlates of integrase targeting? How does the intracellular environment affect the use of certain sequences (or structures) as targets? 2. How does the structure, location, and activity of a region of cellular DNA affect its use as an integration target? In particular, we will test the hypothesis, based on our prior work, that integration has no strong regional preference, and that (contrary to prior ideas), increasing transcriptional activity of a gene does not enhance is use as an integration target, but rather reduces it, as a consequence of interference of bound transcription factors. 3. How does integration affect subsequent expression of the provirus. Why is integration apparently required for efficient expression? What is the role of the integration site in the level of transcription of the integrated provirus? In particular, what is the basis for "position effects" leading to clone-to-clone variation in expression of integrated proviruses?

病毒整合到宿主细胞 DNA 中是逆转录病毒复制的一个核心且独特的特征，并直接导致这一重要病毒组的许多特殊特征，包括它们通过激活和转导癌基因引起癌症的能力、它们的进化持久性宿主种系中的内源性病毒，它们非常广泛的致病谱，以及它们作为基因治疗师载体的能力。 尽管整合过程的生化和结构方面已经得到很好的解决，但许多重要特征仍然未知，包括整合目标的定义、染色质和 DNA 结构和整合特异性之间的关系，以及整合位置的影响。原病毒对其后续表达的影响。 该项目的总体目标是提高我们对病毒与宿主相互作用中整合作用的关键方面的理解。 具体来说，我们将解决以下问题： 1. 我们能否在体外定义整合酶的最佳靶序列（或结构）？ 整合酶靶向的结构相关性是什么？ 细胞内环境如何影响某些序列（或结构）作为靶标的使用？ 2. 细胞 DNA 区域的结构、位置和活性如何影响其作为整合靶点的用途？ 特别是，我们将根据我们之前的工作检验这一假设，即整合没有强烈的区域偏好，并且（与之前的想法相反），增加基因的转录活性并不会增强作为整合目标的使用，而是增强由于结合转录因子的干扰，它会减少。 3.整合如何影响原病毒的后续表达。 为什么有效表达显然需要整合？ 整合位点在整合原病毒的转录水平中起什么作用？ 特别是，导致整合原病毒表达克隆间差异的“位置效应”的基础是什么？