Structural studies of DNA-processing enzymes

DNA 加工酶的结构研究

• 批准号: 9070069
• 负责人: Hideki Aihara
• 金额: $ 53.26万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9070069
• 关键词: Antiviral Agents; Architecture; Area; Chromosomes; Complex; DNA; DNA Damage; DNA-Protein Interaction; Data; Development; Disease; Engineering; Enzymes; Genome; HIV-1; HIV-1 integrase; Human; Integrase; Integrase Inhibitors; Learning; Methods; Natural regeneration; Process; Proteins; Research; Retroviridae; Rous sarcoma virus; Spumavirus; Structure; System; Viral; Virus Integration; X-Ray Crystallography; cellular targeting; design; genome integrity; human disease; improved; inhibitor/antagonist; interest; next generation; novel; novel therapeutics; pathogen; prototype; public health relevance; repaired; structural biology; telomere; tool; transcriptional coactivator p75; viral DNA

 DESCRIPTION (provided by applicant): We will use structural biology tools to investigate DNA-processing enzymes from human pathogens, and those relevant to genome integrity or disease process. An area of particular interest is the mechanism of retrovirus integration by the viral integrase (IN), which forms a multimeric complex with both ends of the linear viral DNA genome and captures a cellular target DNA for concerted insertions of the viral DNA termini. The IN-viral DNA complex formed by HIV-1 IN is the target of a class of antiviral drugs called IN strand-transfer inhibitors. Therefore, a better understanding of the IN-DNA interactions is important for improving the clinically used IN inhibitors as well as designing next-generation inhibitors. We are pursuing crystal structures of IN-DNA complexes from HIV-1 and closely related retroviruses, and structure of those in complex with the host co- factor LEDGF/p75. Our preliminary data suggest that the architectures of the multimeric IN complex containing viral and target DNA molecules are distinct between the extensively characterized prototype foamy virus and other retroviruses with smaller 3-domain INs, including Rous sarcoma virus and HIV-1. Besides retrovirus INs, we are interested in the bacterial and pox viral enzymes responsible for regenerating the hairpin telomeres of linear chromosomes by resolving concatemeric replication intermediates. We also study enzymes involved in cleaning damaged DNA ends for error-free repair by end-joining. Through studies of these systems, we intend to learn novel principles in DNA-protein interactions and obtain structural information that can aid in inhibitor development or design of useful engineered enzymes.

 描述（由申请人提供）：我们将使用结构生物学工具来研究来自人类病原体的 DNA 加工酶，以及与基因组完整性或疾病过程相关的酶，特别感兴趣的领域是病毒整合酶的逆转录病毒整合机制。 (IN)，与线性病毒DNA基因组的两端形成多聚体复合物，并捕获细胞靶DNA以协调插入病毒DNA末端。由HIV-1 IN形成的IN-病毒DNA复合物是a的靶标。班级因此，更好地了解 IN-DNA 相互作用对于改进临床使用的 IN 抑制剂以及设计下一代抑制剂非常重要。 HIV-1 和密切相关的逆转录病毒，以及与宿主辅因子 LEDGF/p75 复合物的结构，我们的初步数据表明，含有病毒和靶 DNA 分子的多聚 IN 复合物的结构与广泛表征的原型泡沫不同。病毒以及其他具有较小 3 结构域 IN 的逆转录病毒，包括 Rous 肉瘤病毒和 HIV-1，除了逆转录病毒 IN 之外，我们还对负责通过解析多联体复制中间体来再生线性染色体发夹端粒的细菌和痘病毒酶感兴趣。研究参与清洁受损 DNA 末端以通过末端连接进行无差错修复的酶，通过对这些系统的研究，我们打算了解 DNA-蛋白质相互作用的新原理并获得可以的结构信息。帮助抑制剂开发或有用的工程酶的设计。