Structural studies of DNA-processing enzymes

DNA 加工酶的结构研究

• 批准号: 9307881
• 负责人: Hideki Aihara
• 金额: $ 61.61万
• 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/9307881
• 关键词: Antiviral Agents; Architecture; Area; Chromosomes; Clinical; Complex; Crystallization; 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通过病毒整合酶（IN）的逆转录病毒整合机制，其与线性病毒DNA基因组的两端形成多聚体复合物并捕获细胞靶DNA用于病毒DNA末端的协同插入。由HIV-1 IN形成的IN-病毒DNA复合物是一类称为IN链转移抑制剂的抗病毒药物的靶点。因此，更好地理解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是重要的。我们正在研究来自HIV-1和密切相关的逆转录病毒的IN-DNA复合物的晶体结构，以及与宿主辅因子LEDGF/p75复合的那些的结构。我们的初步数据表明，架构的多聚体IN复杂的病毒和靶DNA分子之间的广泛表征的原型泡沫病毒和其他逆转录病毒与较小的3-结构域的IN，包括劳斯肉瘤病毒和HIV-1。除了逆转录病毒的IN，我们感兴趣的细菌和痘病毒的酶负责再生的发夹端粒的线性染色体通过解决串联复制中间体。我们还研究了参与清洁受损DNA末端的酶，以便通过末端连接进行无错误修复。通过对这些系统的研究，我们打算学习DNA-蛋白质相互作用的新原理，并获得有助于抑制剂开发或设计有用的工程酶的结构信息。