Chemistry and Biology of Oxidized Purine Lesions in DNA

DNA 中氧化嘌呤损伤的化学和生物学

• 批准号: 7986979
• 负责人: Cynthia J Burrows
• 金额: $ 29.98万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-04 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7986979
• 关键词: 8-hydroxyguanosine; Address; Affect; Affinity; Aging; Allantoin; Amines; Attention; Base Excision Repairs; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Catalysis; Cells; Chemical Structure; Chemicals; Chemistry; Coupled; Crystallography; DNA; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA lesion; DNA-Directed DNA Polymerase; Electrons; Enzymes; Excision; Exposure to; Family; Fapy-dG; Fluorescence; Goals; Guanine; Guanosine; Heart Diseases; Human Development; Hydantoins; Hydrogen Bonding; In Vitro; Infection; Inflammatory Response; Injury; Knockout Mice; Laboratories; Lead; Lesion; Liquid substance; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Modification; Monitor; Mutagenesis; Mutation; Neurologic; Nucleosides; Nucleotide Excision Repair; Oxidants; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Peroxonitrite; Play; Procedures; Process; Proteins; Purines; Relative (related person); Research; Roentgen Rays; Role; Site; Source; Structure; Surface; Testing; Toxic Environmental Substances; Vermont; Work; adduct; aptamer; base; carcinogenesis; design; guanidinohydantoin; human DNA; in vivo; innovation; member; novel; oxidation; public health relevance; purine; repaired; spiroiminodihydantoin; stereochemistry; tumor; 8-hydroxyguanosine; Address; Affect; Affinity; Aging; Allantoin; Amines; Attention; Base Excision Repairs; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Catalysis; Cells; Chemical Structure; Chemicals; Chemistry; Coupled; Crystallography; DNA; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA lesion; DNA-Directed DNA Polymerase; Electrons; Enzymes; Excision; Exposure to; Family; Fapy-dG; Fluorescence; Goals; Guanine; Guanosine; Heart Diseases; Human Development; Hydantoins; Hydrogen Bonding; In Vitro; Infection; Inflammatory Response; Injury; Knockout Mice; Laboratories; Lead; Lesion; Liquid substance; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Modification; Monitor; Mutagenesis; Mutation; Neurologic; Nucleosides; Nucleotide Excision Repair; Oxidants; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Peroxonitrite; Play; Procedures; Process; Proteins; Purines; Relative (related person); Research; Roentgen Rays; Role; Site; Source; Structure; Surface; Testing; Toxic Environmental Substances; Vermont; Work; adduct; aptamer; base; carcinogenesis; design; guanidinohydantoin; human DNA; in vivo; innovation; member; novel; oxidation; public health relevance; purine; repaired; spiroiminodihydantoin; stereochemistry; tumor

DESCRIPTION (provided by applicant): A causative role has been established between compromised DNA repair and the development of human cancers. DNA bases are subject to oxidative stress undergoing chemical modification of guanine and other bases as a result of endogenous oxidants, inflammatory responses to infection and injury, and exposure to redox-active environmental toxins. While much attention has been focused on G to T mutations as a result of the oxidized guanine lesion 8-oxoguanine (OG), this project investigates the hyperoxidized guanine lesions resulting from further oxidation of OG. These lesions constitute a family of highly mutagenic hydantoin structures including spiroiminodihydantoin (Sp) and guanidinohydantoin (Gh) that appear to cause G to C in addition to G to T mutations. In the present work, related members of this class of compounds will be studied to further explore the relationship between oxidation pathway, product structure, and activity with DNA processing enzymes including base excision repair and nucleotide excision repair mechanisms. The new structures to be studied include amine adducts of oxidized guanosine. Novel pathways are proposed that might explain additional observations in the mutagenic spectrum of oxidative stress, including the formation of double-stranded tandem lesions. The specific aims of this work are to: (1) resolve structural questions surrounding the formation of hydantoin lesions in duplex DNA through the use of synthetic lesion-containing oligodeoxynucleotides, mass spectrometry, and x-ray crystallography, (2) understand the roles of base excision repair vs. nucleotide excision repair for hydantoin lesions via in vitro and in vivo biochemical assays with a particular focus on the role of the Nei-like (hNEIL1) and Fpg glycosylases, and (3) develop innovative assays to detect hydantoin products in cell lysates by generating aptamers to DNA lesions coupled to fluorescent readouts. PUBLIC HEALTH RELEVANCE: Oxidative stress plays a significant role in the damaging DNA creating lesions that underlie cancer, aging, neurological and cardiac disorders. This project will help define how specific oxidized DNA bases and base adducts lead to mutations related to cancer. In addition, methods will be developed to monitor the formation and excision of these damaged bases from DNA, revealing new information about the relationship between DNA damage and cancer.

描述（由申请人提供）：在DNA修复和人类癌症的发展之间已经确定了致病作用。由于内源性氧化剂，对感染和损伤的炎症反应以及暴露于氧化还原活性的环境毒素，DNA碱受到鸟嘌呤和其他碱的化学修饰的氧化应激。尽管由于氧化的鸟嘌呤病变8-氧气苷（OG）的氧化性鸟嘌呤病变（OG），该项目研究了GOM的G突变，但该项目研究了由OG的进一步氧化而导致的高氧化鸟嘌呤病变。这些病变构成了一个高度诱变的羟基毒素结构的家族，包括螺旋氨基磷酸（SP）和瓜尼迪诺羟多托（GH），这些家族似乎使G到C到T突变外似乎会导致G到C。在目前的工作中，将研究此类化合物的相关成员，以进一步探索氧化途径，产品结构和活性与DNA加工酶之间的关系，包括碱基切除修复和核苷酸切除修复机制。要研究的新结构包括氧化鸟嘌呤的胺加合物。提出了新的途径，可以解释氧化应激的诱变光谱中的其他观察结果，包括形成双链串联病变。这项工作的具体目的是：（1）通过使用含有合成病变的寡脱氧核苷酸，质谱法和X射线结晶学的含有合成病变的寡脱氧核苷酸，（2）了解基本切除维修效果的效果，解决了双层DNA中综合性病变的结构问题。生化测定特别关注Nei样（HNEIL1）和FPG糖基酶的作用，以及（3）通过生成适合于DNA病变与荧光读数的DNA病变，开发创新的测定方法，以检测细胞裂解物中的羟托斯产物。 公共卫生相关性：氧化应激在破坏性DNA中起着重要作用，从而产生癌症，衰老，神经系统疾病和心脏疾病的病变。该项目将有助于定义特定的氧化DNA碱基和碱加合物如何导致与癌症相关的突变。此外，将开发方法来监测DNA受损碱的形成和切除，从而揭示有关DNA损伤与癌症之间关系的新信息。