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（由适用提供）：我们将使用结构生物学工具来研究人类病原体的DNA处理酶，以及与基因组完整性或疾病过程相关的酶。特别感兴趣的区域是病毒整合酶（IN）逆转录病毒整合的机理，该机制形成了一个多聚体复合物，具有线性病毒DNA基因组的两端，并捕获了一个细胞靶DNA，以协调病毒DNA末端的插入。由HIV-1形成的病毒内DNA复合物是在链转移抑制剂中称为一类抗病毒药的靶标。因此，对DNA相互作用的更好理解对于改善抑制剂和设计下一代抑制剂的临床使用至关重要。我们正在从HIV-1和密切相关的逆转录病毒中追求DNA复合物的晶体结构，以及与宿主共同因素LEDGF/p75相关的结构。我们的初步数据表明，含有病毒和靶DNA分子的复合物中倍数的体系结构在广泛特征的原型泡沫病毒和其他具有较小三域INS的逆转录病毒之间与众不同，包括Rous肉瘤病毒和HIV-1。除了逆转录病毒INS外，我们对通过解决串联复制中间体来再生线性染色体的发夹端粒的细菌和痘病毒酶感兴趣。我们还研究涉及清洁受损DNA末端的酶，以通过最终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，并获得可以帮助抑制剂开发或设计有用的工程酶的结构信息。