STRUCTURAL IDENTIFICATION AND CHARACTERIZATION OF A NEW DNA PHOTOPRODUCT

新型 DNA 照片产品的结构鉴定和表征

• 批准号: 8168731
• 负责人: JOHN-STEPHEN Adolfino TAYLOR
• 金额: $ 1.28万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-10 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168731
• 关键词: Attention; Computer Retrieval of Information on Scientific Projects Database; Cytosine; DNA biosynthesis; Deamination; Funding; Goals; Grant; High Pressure Liquid Chromatography; Institution; Kinetics; Link; Mass Spectrum Analysis; Measures; Preparation; Pyrimidine Dimers; Research; Research Personnel; Resources; Site; Skin Cancer; Source; Structure; Structure-Activity Relationship; Techniques; Time; Ultraviolet B Radiation; United States National Institutes of Health; X-Ray Crystallography; design; dimer; irradiation; sunlight-induced; transition mutation; ultraviolet irradiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. DNA synthesis past deamination products of cytosine and 5-methylcytosine-containing cis-syn pyrimidine dimer photoproducts may be one of the principal mechanisms by which sunlight induces C-to-T transition mutations linked to skin cancer. To understand better the structure-activity relationships governing photoproduct deamination, we designed the ODN sequence d(GTATCATGAGGTGC) containing a unique TC site to enable preparation of a dT[c,s]C dimer by UV irradiation to measure the kinetics of deamination of cytosine photoproducts. Instead of the expected T[c,s]C photoproduct, an abundant unknown with an unusually short HPLC retention time attracted our attention during the course of UVB irradiation (312 nm) of d(GTATCATGAGGTGC). This product only appeared at low pH (below 6). Our goal is to elucidate the structure of this unknown major photoproduct by combining various techniques such as HPLC, mass spectrometry, NMR, and X-Ray crystallography.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 DNA合成过去的脱氨基产物的胞嘧啶和5-甲基胞嘧啶的顺式-syn嘧啶二聚体的光产物可能是其中一个主要的机制，阳光诱导的C到T的转换突变与皮肤癌。 为了更好地理解光产物脱氨基的结构-活性关系，我们设计了含有独特TC位点的ODN序列d（GTATCATGAGGTGC），以使得能够通过UV照射制备dT[c，s]C二聚体来测量胞嘧啶光产物脱氨基的动力学。 而不是预期的T[c，s]C的光产物，一个丰富的未知的异常短的HPLC保留时间吸引了我们的注意力在UVB照射（312 nm）的过程中的d（GTATCATGAGGTGC）。该产物仅在低pH（低于6）下出现。我们的目标是通过结合各种技术，如HPLC，质谱，NMR和X射线晶体学来阐明这种未知的主要光产物的结构。