UV DAMAGE AND REPAIR AT THE DNA SEQUENCE LEVEL

DNA 序列水平的紫外线损伤和修复

• 批准号: 2154869
• 负责人: Gerd P Pfeifer
• 金额: $ 18.71万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2154869
• 关键词: 5 methylcytosine; DNA binding protein; DNA damage; DNA footprinting; DNA repair; amination; chromatin; cytosine; dimer; fibroblasts; gene mutation; genetic disorder; genetic mapping; genetic transcription; human genetic material tag; human tissue; hypoxanthine phosphoribosyltransferase; molecular site; nucleic acid sequence; polymerase chain reaction; protooncogene; skin neoplasms; ultraviolet radiation

The incidence of skin cancer, the most common type of human cancer, is increasing at an alarming rate in the United States and worldwide. The damage of DNA bases by ultraviolet (UV) radiation causes mutations and UV light is strongly implicated in the development of human basal and squamous cell carcinoma as well as the less common but more lethal melanoma. To understand UV carcinogenesis, a more detailed knowledge of the molecular mechanisms of UV damage formation, DNA repair processes and mutation induction is necessary. We will apply uniquely sensitive, mostly PCR-based techniques, to map UV-induced lesions and their repair rates as well as the mutations they produce at the DNA sequence level in human genes. We propose to investigate the molecular mechanisms of selective UV damage formation in vivo, and to analyze DNA sequence-specific repair rates in several human genes. Possible molecular mechanisms of sequence- specific repair of UV-induced lesions will be elucidated by comparative analysis of chromatin structure and protein-DNA interactions at sequences showing differential repair rates. Variations of the ligation-mediated PCR technique will be used to search for in vivo processing intermediates of UV photoproducts. We will attempt to develop new methodology to study repair of (6-4) photoproducts at the DNA sequence level. The relationship between high UV damage frequency, slow repair rates and mutagenesis at specific sites will be determined by using an allele-specific PCR assay for-mutation detection. Finally, we will investigate whether genetic defects in human DNA repair deficiency and cancer-prone syndromes are related to alterations in sequence-specific DNA repair.

皮肤癌的发病率是人类癌的最常见类型，是 在美国和全球以惊人的速度提高。这 通过紫外线（UV）辐射损害DNA碱基的损伤会导致突变和紫外线 光与人类基础和 鳞状细胞癌以及不太常见但更致命的 黑色素瘤。为了了解紫外线发生，更详细地了解 紫外线损伤形成，DNA修复过程的分子机制和 突变诱导是必要的。我们将采用独特的敏感，主要是 基于PCR的技术，以将紫外线诱导的病变及其维修率映射为 以及它们在人类DNA序列水平上产生的突变 基因。我们建议研究选择性紫外线的分子机制 体内损伤形成，并分析DNA序列特异性修复 几个人类基因的速率。序列可能的分子机制 紫外线诱导的病变的特定修复将通过比较阐明 分析序列上染色质结构和蛋白-DNA相互作用 显示差异维修率。连接介导的PCR的变化 技术将用于搜索体内处理中间体 紫外光产物。我们将尝试开发新的方法来研究 在DNA序列水平上修复（6-4）光产物。关系 在高紫外线损伤频率，缓慢的维修率和诱变之间 特定站点将通过使用等位基因特异性PCR测定法确定 福音检测。最后，我们将研究遗传是否 人DNA修复缺乏和容易发生癌症的缺陷是 与序列特异性DNA修复的改变有关。