SPECIFICITY OF DNA/RNA BINDING BY A ZINC FINGER PROTEIN

锌指蛋白结合 DNA/RNA 的特异性

• 批准号: 2608940
• 负责人: JOEL M. GOTTESFELD
• 金额: $ 22.42万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2608940
• 关键词: DNA; RNA; X ray crystallography; chemical binding; nuclear magnetic resonance spectroscopy; nucleic acid sequence; nucleic acid structure; protein structure; site directed mutagenesis; transcription factor

DESCRIPTION: TFIIIA binds with high affinity to the 5SrRNA gene as a specific transcription factor and to 5S RNA in a 7S RNP storage particle. This proposal focuses on both the DNA and RNA sequence elements required for high affinity interactions with TFIIIA and the roles played by individual Zn finger domains in specifying high affinity DNA and RNA binding. Different sets of Zn fingers are required for high affinity DNA and RNA binding. Experiments are proposed to investigate the structural elements in DNA and RNA recognized by TFIIIA as well as further investigation into the role of specific amino acids (within specific fingers and in the linker region between fingers). Mutagenesis phage display will be used to investigate the amino acid differences that distinguish RNA and DNA specific Zn fingers. Filter binding and fluorescent anisotrophy solution methods will be used to determine the energetic processes that drive binding reactions to DNA and RNA. Continued collaborative efforts will attempt to obtain high resolution structures of Zn finger DNA and RNA complexes by heteronuclear NMR methods and X-ray crystallography.

描述：TFIIIA以高亲和力与5SrRNA基因结合， 特异性转录因子和7S RNP储存颗粒中的5S RNA。 这项建议集中在DNA和RNA序列所需的元素， 与TFIIIA的高亲和力相互作用以及单个Zn 指结构域在指定高亲和力DNA和RNA结合中的作用。 不同 高亲和力DNA和RNA结合需要锌指组。 提出了研究DNA中结构元件的实验， TFIIIA识别的RNA以及进一步研究 特定的氨基酸（在特定的指内和接头区中 手指之间）。 突变噬菌体展示将用于研究 区别RNA和DNA特异性锌指的氨基酸差异。 将使用滤膜结合法和荧光各向异性溶液法， 确定驱动DNA结合反应的能量过程， 核糖核酸 持续的合作努力将试图获得高分辨率 锌指DNA和RNA复合物结构的核磁共振研究 和X射线晶体学