Sequence Effects of Arylamine-DNA Adducts: Repair and Replication

芳胺-DNA 加合物的序列效应：修复和复制

• 批准号: 7622808
• 负责人: Bongsup P Cho
• 金额: $ 29.99万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7622808
• 关键词: Abbreviations; Accounting; Affinity; Aromatic Amines; Binding; Cancer Etiology; Carcinogens; Cells; Circular Dichroism; Classification; Complex; Condition; DNA; DNA Adduction; DNA Adducts; DNA Damage; DNA Polymerase I; DNA-Protein Interaction; Data; Deletion Mutation; Development; Differential Scanning Calorimetry; EMSA; Electrostatics; Entropy; Environment; Equilibrium; Escherichia coli; Exonuclease; Fluorescence Spectroscopy; Fluorine; Funding; Gel; Goals; Heterogeneity; Hot Spot; Human; Induced Mutation; Investigation; Knowledge; Lesion; Link; Location; Malignant Neoplasms; Methodology; Methods; Molecular; Molecular Conformation; Molecular Probes; Mutagenesis; Mutation; Nuclear Magnetic Resonance; Nucleotide Excision Repair; Nucleotides; Outcome; Pathway interactions; Physiological; Plasma; Play; Polymerase; Population; Prevention; Procedures; Process; Property; Proteins; Range; Replication-Associated Process; Research; Risk Assessment; Role; Scanning; Simulate; Specificity; Structure; Surface; Surface Plasmon Resonance; Surgical incisions; System; Testing; Thermodynamics; Work; XPA gene; abstracting; acetylaminofluorene; adduct; base; chemical carcinogenesis; conformer; design; enthalpy; grasp; human tissue; innovation; insight; programs; prototype; repaired; research study; size; structural biology; theories; tool

Project Summary/Abstract The long term goal of our research program is to elucidate the molecular mechanisms of DNA adduct-induced chemical carcinogenesis. Aromatic amines are well-known environmental human carcinogens. In particular, arylamine-DNA adduct formation has been confirmed in various human tissues and is believed to induce mutation. We have previously shown that arylamine adducts in DNA exist in three well-defined conformations: stacked (S), external B-type (B), and wedge (W). The conformation depends on the location of the carcinogen moiety in the DNA molecule, and the population ratios of the types under physiological conditions are sequence-dependent. In this application, we hypothesize that arylamine-induced repair and mutation is conformation (S/B/W) specific. We propose four major aims to help define adduct conformation and examine their specific effects on repair (initial damage recognition) and replication (replication fork heterogeneity). Specifically, these aims focus on: (1) conformation-specific repair in a human nucleotide excision repair (NER) system, (2) long-range sequence effects, (3) the thermodynamics of sequencedependent slippage-induced frameshift mutagenesis, and (4) replication fork conformational heterogeneity. We will employ not only existing dynamic 19F NMR/CD, EMSA and fluorescence spectroscopy, but also innovative chip-based surface plasma resonance (SPR) and differential scanning calorimetric (DSC) procedures, creating a powerful suite of biophysical methodologies. Successful completion of the proposed aims will help us gain a better grasp on the protein-DNA interactions involved in human NER and trans-lesion synthesis, which have important implications for resolving the molecular details of cancer etiology. Such knowledge will also be of help in the development of sensible prevention and risk assessment strategies.

项目概要/摘要 我们研究计划的长期目标是阐明 DNA 的分子机制 加合物诱导的化学致癌作用。芳香胺是众所周知的环境人类胺 致癌物质。特别是，芳基胺-DNA 加合物的形成已在多种人类体内得到证实。 组织并被认为会诱导突变。我们之前已经证明 DNA 中的芳基胺加合物 存在三种明确的构象：堆叠（S）、外部B型（B）和楔形（W）。这 构象取决于致癌物部分在DNA分子中的位置，以及 生理条件下各类型的种群比例取决于序列。在这个 应用中，我们假设芳胺诱导的修复和突变是构象（S/B/W） 具体的。我们提出了四个主要目标来帮助定义加合物构象并检查其具体 对修复（初始损伤识别）和复制（复制叉异质性）的影响。 具体来说，这些目标集中在：（1）人类核苷酸切除中的构象特异性修复 修复（NER）系统，（2）长程序列效应，（3）序列依赖的热力学 滑移诱导的移码突变，以及（4）复制叉构象 异质性。我们不仅会采用现有的动态 19F NMR/CD、EMSA 和荧光 光谱，还包括创新的基于芯片的表面等离子体共振 (SPR) 和微分 扫描量热 (DSC) 程序，创建了一套强大的生物物理方法。 成功完成所提出的目标将有助于我们更好地掌握蛋白质-DNA 人类 NER 和跨损伤合成中涉及的相互作用，这对 解决癌症病因学的分子细节。这些知识也将有助于 制定明智的预防和风险评估策略。