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: 8022957
• 负责人: Bongsup P Cho
• 金额: $ 28.88万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8022957
• 关键词: Accounting; Affinity; Aromatic Amines; Binding; Cancer Etiology; Carcinogens; Cells; Complex; 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; Etiology; Fluorescence Spectroscopy; Funding; Gel; Goals; Health; Heterogeneity; Hot Spot; Human; Induced Mutation; Investigation; Kinetics; Knowledge; Lesion; Link; Location; Malignant Neoplasms; Methodology; Methods; Molecular; Molecular Conformation; Molecular Probes; Mutagenesis; Mutation; Nucleotide Excision Repair; Outcome; Pathway interactions; Physiological; Plasma; Play; Polymerase; Population; Prevention; Primer Extension; Procedures; Process; Property; Proteins; Replication-Associated Process; Research; Risk Assessment; Role; Scanning; Simulate; Specificity; Structure; Surface; Surface Plasmon Resonance; Surgical incisions; System; Testing; Thermodynamics; Work; XPA gene; adduct; adverse outcome; base; cancer initiation; chemical carcinogenesis; conformer; design; enthalpy; environment related cancer; environmental chemical; grasp; human tissue; in vivo; innovation; insight; programs; prototype; public health relevance; repaired; research study; structural biology; theories; tool

DESCRIPTION (provided by applicant): 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-specific (S, B, W). 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 and long-range sequence effects, (2) damage recognition (protein-DNA interaction) (3) the thermodynamics of sequence-dependent slippage-induced frameshift mutagenesis, and (4) replication fork conformational heterogeneity and polymerase binding. 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. PUBLIC HEALTH RELEVANCE: The primary causes of sporadic human cancers are environmental. Aromatic amines are among the most notorious environmental chemicals that are implicated in the etiology of human cancers. Formation of arylamine-DNA adducts has been confirmed in various human tissues and is believed to induce chemical carcinogenesis. Hence it is imperative to elucidate how these lesions are repaired and replicated in vivo at the atomic and molecular levels. The key molecular players (adduct structures, polymerases, repair proteins) producing adverse outcomes must be identified, characterized, and understood in order to devise appropriate prevention and risk assessment strategies.

描述（由申请人提供）：我们研究计划的长期目标是阐明DNA加合物诱导的化学癌变的分子机制。芳香胺是著名的环境人类致癌物。特别是，在各种人体组织中已经证实了芳胺-DNA加合物的形成，并被认为会诱导突变。我们先前已经表明，DNA中的芳胺加合物存在于三种明确的构象中：堆叠（S），外部B型（B）和楔形（W）。构象取决于致癌物在DNA分子中的位置，生理条件下类型的种群比率依赖于序列。在此应用中，我们假设芳基胺诱导的修复和突变是构象特异性的（S，B，W）。我们提出了四个主要旨在帮助定义加合构象并检查其对修复（初始损害识别）和复制（复制叉异质性）的特定影响的主要目标。 Specifically, these aims focus on: (1) conformation-specific repair in a human nucleotide excision repair (NER) system and long-range sequence effects, (2) damage recognition (protein-DNA interaction) (3) the thermodynamics of sequence-dependent slippage-induced frameshift mutagenesis, and (4) replication fork conformational heterogeneity and polymerase binding.我们将不仅采用现有的动态19F NMR/CD，EMSA和荧光光谱，还将采用创新的基于创新的基于芯片的表面等离子体共振（SPR）和差异扫描量热量法（DSC）程序，从而创建了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了解人类NER和跨性质合成所涉及的蛋白质-DNA相互作用，这对解决癌症病因的分子细节具有重要意义。这些知识也将有助于开发明智的预防和风险评估策略。 公共卫生相关性：零星人类癌症的主要原因是环境的。芳香胺是与人类癌症的病因有关的最臭名昭著的环境化学物质之一。在各种人体组织中已经确认了芳胺-DNA加合物的形成，并被认为会诱导化学癌变。因此，必须阐明如何在原子和分子水平的体内修复和复制这些病变。必须确定，表征和理解产生不良结果的关键分子参与者（加合结构，聚合酶，修复蛋白），以设计适当的预防和风险评估策略。