Structure and Function of DNA Repair Enzymes and Cancer

DNA 修复酶的结构和功能与癌症

• 批准号: 10014581
• 负责人: Sylvie Doublie
• 金额: $ 183.72万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-03 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10014581
• 关键词: Affect; Base Excision Repairs; Basic Science; Biochemical; Biochemistry; Bioinformatics; Biological; Cancer Biology; Cancer Etiology; Cell Culture Techniques; Cells; Cellular biology; Chromatin; Chromatin Structure; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA analysis; DNA glycosylase; Data; Defect; Enzymes; Failure; Family; Genes; Genetic Variation; Genomic Instability; Germ-Line Mutation; Goals; Human; Human Cloning; Human Genetics; Individual; Knowledge; Lead; Malignant Neoplasms; Mutation; Pathway interactions; Predisposition; Process; Proteins; Resistance; Resource Sharing; Risk Assessment; Structure; Testing; Translating; Treatment Efficacy; Variant; Work; cancer prevention; cancer therapy; carcinogenesis; gene repair; insight; molecular imaging; multidisciplinary; mutant; personalized medicine; predictive marker; programs; repair enzyme; repaired; response; single molecule; structural biology; targeted treatment; tumor; tumor progression

PROJECT SUMMARY On a daily basis, each cell in our body is bombarded with some 30,000 endogenous DNA damages per day, the vast majority of which are repaired by Base Excision Repair (BER). The central hypothesis of this Program is that, understandably, defects in this BER process drive human carcinogenesis and affect responses to cancer treatments. The overall goal of this Program Project is to functionally characterize human genetic variation in the BER enzymes. To accomplish this, we are characterizing potentially damaging germline and tumor-associated SNPs in the B E R DNA glycosylases, and a number of the downstream enzymes in the BER pathway, using our strengths in bioinformatics, cell biology, biochemistry, structural biology and single molecule imaging in order to determine the functions of the wild-type and variant proteins. Our preliminary data suggest that fundamental mechanistic studies are essential for interpreting human genetic variation and its influence on cancer etiology and tumor progression. Our program is informed and driven by the identification and characterization of germline and tumor-associated enzyme variants that may contribute to the altered DNA repair capacity of human BER enzymes. To realize our goals, variants in the BER genes are prioritized for study using bioinformatics (Core A) as well as enzymatic activity and structural information (Project 2). We then test for functional consequences of the variation using a powerful combination of biological, biochemical, structural, and single-molecule approaches. Core A provides the bioinformatics underpinning of the Program; Project 1, the biological studies of the human variant proteins in human cells; Project 2, the structure/function and biochemical analyses of the BER glycosylases; Project 3, the study of the BER repair process in chromatin; and Project 4, insights into the damage target search of the wild-type and variant BER proteins. Core B provides purified proteins to Projects 2-4 and cell cultures of variant clones to Projects 1-4, while Core C provides the administrative support. Taken together, the mechanistic results obtained by this Program Project provide a unique opportunity to underpin critical questions surrounding cancer etiology and cancer treatment, thus substantially impacting personalized medicine.

项目总结 每天，我们体内的每个细胞每天都会受到大约30,000个内源性DNA损伤的轰炸， 其中绝大多数是通过碱基切除修复(BER)修复的。这一计划的中心假设 可以理解的是，BER过程中的缺陷会推动人类癌症的发生，并影响对 癌症治疗。该计划项目的总体目标是从功能上描述人类基因 误码率酶的变异。为了实现这一点，我们正在鉴定潜在的破坏性生殖系和 B、E、R DNA糖基酶中与肿瘤相关的SNPs，以及在B、E、R基因中的一些下游酶 BER途径，利用我们在生物信息学、细胞生物学、生物化学、结构生物学和单细胞 分子成像以确定野生型和变异蛋白的功能。我们的 初步数据表明，基本的机械学研究对于解释人类是必不可少的 基因变异及其对肿瘤病因和肿瘤进展的影响。我们的节目是知情的， 由生殖系和肿瘤相关酶变异体的鉴定和特征驱动 可能有助于人类BER酶DNA修复能力的改变。为了实现我们的目标， BER基因优先用于生物信息学(核心A)以及酶活性和 结构信息(项目2)。然后，我们使用一个强大的 生物、生化、结构和单分子方法的结合。核心A提供 该计划的生物信息学基础；项目1，人类变异蛋白的生物学研究 人类细胞；项目2，BER糖基酶的结构/功能和生化分析；项目3， 染色质中误码修复过程的研究；以及项目4，对损伤目标搜索的洞察 野生型和变异型BER蛋白。核心B为项目2-4和细胞提供纯化的蛋白质 项目1-4的不同克隆的文化，而核心C提供行政支持。加在一起， 该计划项目所取得的机械性成果提供了一个独特的机会来支持关键的 围绕癌症病因和癌症治疗的问题，从而极大地影响个性化 医药。