SYNTHESIS & STRUCTURE OF OLIGODEOXYNUCLEOTIDES WITH DEFINED DNA DAMAGE

合成

• 批准号: 6575676
• 负责人: FRANCIS JOHNSON
• 金额: $ 21.9万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6575676
• 关键词: DNA damage; DNA repair; adduct; alkylation; chemical carcinogen; environmental toxicology; mutagens; nuclear magnetic resonance spectroscopy; nucleic acid chemical synthesis; nucleic acid sequence; oligonucleotides; phenolic amine; site directed mutagenesis; synthetic nucleic acid

The synthetic organic chemistry described in this section forms a basis for the research projects described by other members of the Program. A fundamental objective is the synthesis of a series of modified DNA nucleosides that will be incorporated site-specifically into oligonucleotides using automated, solid-state methods. The modified deoxynucleosides selected for study fall into two classes. The first group is related to our investigations on oxidative DNA damage. These substances will be used (a) to construct affinity chromatography substrates for the purification and isolation of DNA repair enzymes (MutM and MutY proteins) that deal with oxidative lesions and (b) to study the mechanisms of action of these enzymes (principally MutY) in the repair process. The second group of deoxynucleosides have modifications related to carcinogenic amines and nitro compounds. They are subdivided into two classes: (a) those that relate to C8-dG adducts and (b) those that rise by attack of the ultimate carcinogen on the N2-position of deoxyguanosine. These defined adducts will be used to establish the mutagenic spectrum generated by the original carcinogen. In addition, they should aid in the solution of structural problems associated both with the nucleosides themselves and with oligomeric DNA containing them. For these purposes, 13C and 15N-labeled nucleosides will be synthesized. Research on the use of new protecting groups for DNA synthesis also will be pursued. This should allow otherwise difficult post-synthetic introduction of base-sensitive lesions into DNA. Analytical studies will investigate the formation of new oxidation products, including cross- linked deoxynucleoside dimers whose structure has not been characterized, and will establish accurate, sensitive assays for oxidation products derived from nuclear and mitochondrial DNA.

本节中描述的合成有机化学形成了基础 对于该计划的其他成员描述的研究项目。一个 基本目标是一系列修饰的DNA的合成 核苷将特定于现场纳入 使用自动固态方法的寡核苷酸。修改后 选择进行研究的脱氧核苷分为两类。第一个 组与我们对氧化DNA损伤的研究有关。这些 物质将用于（a）构建亲和力色谱法 纯化和分离DNA修复酶的底物（MUTM 和混合蛋白）处理氧化病变，并且（b）研究 这些酶的作用机制（主要是模仿）在修复中 过程。第二组脱氧核苷与修改有关 致癌胺和硝基化合物。他们被细分为两个 课程：（a）与C8-DG加合物相关的和（b）那些相关的人 通过对N2位置的最终致癌物的攻击 脱氧鸟苷。这些定义的加合物将用于建立 原始致癌物产生的诱变谱。此外， 他们应该有助于解决相关的结构性问题 与核苷本身以及含有核苷的DNA一起使用。 为了这些目的，将合成13C和15N标记的核苷。 关于使用新保护组进行DNA合成的研究也将 被追捕。这应该允许否则困难的合成后 将碱敏感病变引入DNA。分析研究将 研究新氧化产物的形成，包括跨 连接的脱氧核苷二聚体，其结构尚未表征， 并将为氧化产物建立准确，敏感的测定法 源自核和线粒体DNA。