BIOLOGICAL CONSEQUENCES OF SITE-SPECIFIC DAMAGE TO DNA

DNA 特定位点损伤的生物学后果

• 批准号: 2090283
• 负责人: Louis J Romano
• 金额: $ 14.89万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-05-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2090283
• 关键词: DNA damage; DNA repair; adduct; carbopolycyclic compound; chemical carcinogen; chemical structure function; conformation; cyclic amine; helicase; microorganism genetics; nuclear magnetic resonance spectroscopy; nucleic acid chemical synthesis; nucleic acid sequence; nucleic acid structure; oligonucleotides; site directed mutagenesis

The goals of this proposal are to continue the development of a general method for the preparation of oligodeoxyribonucleotides containing chemically well-defined damage at unique and specific locations, to use these molecules to determine how specific adducts affect the three- dimensional structure of a DNA duplex, and at attempt to relate these structural changes to the mutations that they induce. The data generated in these studies will be used to test the hypothesis that the type of mutation induced is dependent on the adduct structure and the DNA sequence within which it is located. We will also attempt to specifically relate this analysis to the question of the importance of non-targeted or semi-targeted mutagenesis. In addition, these site- specifically modified templates will also be used to characterize the stable complex which we have observed to form between a DNA lesion and a DNA helicase. This proposal is focussed on two important and well- studied classes of carcinogenic DNA adducts: aromatic amines and polycyclic aromatic hydrocarbons. However, our long-term objective is to develop techniques to place any DNA lesion into a specific DNA sequence, even including those where the chemistry needed to prepare the lesion is not now known or where the instability of the lesion to currently available oligonucleotide synthesis technology makes the synthesis prohibitive. The relevance of this work towards the progress of understanding the initiation of cancer in eukaryotic cells is two- fold. First, studying the relationship between adduct structure within a specific sequence context and induced mutations and mutation frequency will lead to an understanding of the parameters which define a mutational hotspot, many of which are thought to be important in oncogene activation. Second, studying the irreversible binding of a protein to a DNA adduct site may clarify the role these structures might play in the induction of chromosomal breaks or rearrangements.

该提案的目标是继续开发通用的 含有寡脱氧核糖核苷酸的制备方法 在独特和特定位置进行化学明确的损伤，以使用 这些分子以确定特定加合物如何影响三- DNA双链体的维度结构，并试图将这些联系起来 它们诱导的突变的结构变化。 生成的数据 在这些研究中将用于检验以下假设： 诱导的突变取决于加合物结构和 DNA 它所在的序列。 我们也将尝试 具体将此分析与重要性问题联系起来 非靶向或半靶向诱变。 此外，这些网站—— 特别修改的模板也将用于表征 我们观察到 DNA 损伤和 DNA 损伤之间形成的稳定复合物 DNA解旋酶。 该提案集中于两个重要且良好的方面： 研究了一类致癌 DNA 加合物：芳香胺和 多环芳烃。 然而，我们的长期目标是 开发将任何 DNA 损伤置于特定 DNA 中的技术 序列，甚至包括制备所需化学物质的序列 目前尚不清楚病变部位或病变部位的不稳定性 目前可用的寡核苷酸合成技术使得 合成禁止。 这项工作与进展的相关性 了解真核细胞癌症发生的关键有两个： 折叠。 首先，研究内加合物结构之间的关系 特定的序列背景和诱导的突变和突变频率 将导致对定义突变的参数的理解 热点，其中许多被认为在癌基因中很重要 激活。 其次，研究蛋白质与蛋白质的不可逆结合。 DNA 加合物位点可以阐明这些结构在 诱导染色体断裂或重排。