PROPERTIES OF A NEW HUMAN DNA REPAIR COMPLEX

新型人类 DNA 修复复合物的特性

• 批准号: 2608947
• 负责人: ANDREW EISEN
• 金额: $ 9.81万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2608947
• 关键词: ADP ribosylation; DNA binding protein; DNA directed DNA polymerase; DNA repair; SDS polyacrylamide gel electrophoresis; chromosome translocation; computer assisted sequence analysis; enzyme activity; enzyme complex; genetic disorder; genetic recombination; high performance liquid chromatography; human genetic material tag; immunoaffinity chromatography; immunoprecipitation; intermolecular interaction; laboratory rabbit; molecular cloning; neoplastic transformation; nucleic acid sequence; oncogenes; phosphoester ligase; polymerase chain reaction; protein biosynthesis; ADP ribosylation; DNA binding protein; DNA directed DNA polymerase; DNA repair; SDS polyacrylamide gel electrophoresis; chromosome translocation; computer assisted sequence analysis; enzyme activity; enzyme complex; genetic disorder; genetic recombination; high performance liquid chromatography; human genetic material tag; immunoaffinity chromatography; immunoprecipitation; intermolecular interaction; laboratory rabbit; molecular cloning; neoplastic transformation; nucleic acid sequence; oncogenes; phosphoester ligase; polymerase chain reaction; protein biosynthesis

The balance between damage and repair of genomic DNA is vital to both the pathogenesis and the treatment of malignancies. When damage to genomic DNA exceeds the ability to repair that damage the consequences can be mutagenic, carcinogenic or even lethal. This is clearly seen in individuals with genetic deficiencies in DNA repair processes such as Bloom's and other syndromes. These individuals have a high predilection for developing chromosomal abnormalities associated with a variety of cancers. Conversely there is also a benefit to be gained when damage to genomic DNA exceeds the ability to repair that damage. Lethal damage is imposed upon genomic DNA by many chemotherapeutic agents. Thus, the enzymes of DNA recombination and repair are especially useful targets for primary and adjunctive chemotherapy. Further development of these enzymes as targets for chemotherapy will depend upon a more complete understanding of the DNA ligand recognized by these enzymes and how various enzymes co-ordinately act to repair damaged DNA. Some of these enzymes probably act upon DNA alone. However, it has become clear that many proteins that bind to, modify or regulate DNA function do so as part of multi-protein complexes. It will be quite important to identify the DNA repair enzymes that associate into complexes for the rational design of ligands that can serve as chemotherapeutic agents. This laboratory has recently identified the formation of a human DNA repair complex that assembles on a unique DNA ligand. A trio of human nuclear proteins binds to the recombinogenic Chi (5'-GCTGGTGG-3') sequence and was affinity purified in a single step. The trio contains a Chi- binding protein and two associated proteins. The Chi binding species was identified as the DNA repair enzyme poly (ADP-ribose) polymerase. The second species forms a stable adenylate adduct and its electrophoretic mobility suggests it is DNA ligase l. The third species co-sediments with the ligase and is probably a newly described "inhibitor" of ligase. Under defined conditions the complex acts like a Chi sequence-specific recombinase. This proposal seeks to further characterize the biochemical and biological activities of this new human DNA repair complex. The complex may be involved in the pathogenesis of oncogenic chromosomal translocations and serve as a target for a new class of chemotherapeutic drugs.

损害和修复基因组DNA之间的平衡对于这两者都至关重要 发病机理和恶性肿瘤的治疗。当损害基因组DNA 超过修复损害后果的能力可能是 诱变，致癌甚至致命。这清楚地看到了 在DNA修复过程中患有遗传缺陷的个体，例如 布卢姆和其他综合症。 这些人有很高的偏好 用于开发与各种相关的染色体异常 癌症。相反，当损害损坏时，也有一个好处 基因组DNA超过修复损伤的能力。致命伤害是 许多化学治疗剂对基因组DNA施加。 因此，DNA重组和修复的酶特别有用 初级和辅助化疗的靶标。进一步发展 这些酶作为化学疗法的靶标将取决于更完整的 对这些酶识别的DNA配体的理解以及如何 各种酶协同作用以修复受损的DNA。其中一些 酶可能仅作用于DNA。但是，很明显 许多结合，修改或调节DNA功能的蛋白质确实如此 多蛋白质复合物。识别DNA非常重要 维修酶，将其关联为复合物进行合理设计 可以用作化学治疗剂的配体。 该实验室最近确定了人类DNA的形成 修复在独特的DNA配体上组装的复合物。三人 核蛋白与重组CHI（5'-GCTGGTGG-3'）序列结合 并在一个步骤中纯化了亲和力。三人包含一个 结合蛋白和两个相关蛋白。 Chi结合物种是 被鉴定为DNA修复酶聚（ADP-核糖）聚合酶。这 第二种形成稳定的腺苷加合物及其电泳 迁移率表明它是DNA连接酶L。第三个物种与 连接酶，可能是新描述的连接酶的“抑制剂”。在下面 定义条件该复合物的作用像是CHI序列特异性的 重组酶。该建议旨在进一步描述生化 和这种新的人类DNA修复复合物的生物学活性。这 复合物可能参与致癌染色体的发病机理 易位并作为新的化学治疗类别的目标 毒品。