STRUCTURAL STUDIES OF TRANSCRIPTIONAL ACTIVATION

转录激活的结构研究

• 批准号: 2634836
• 负责人: Rachel E Klevit
• 金额: $ 16.14万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2634836
• 关键词: DNA binding protein; SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; fungal genetics; gene expression; genetic transcription; high performance liquid chromatography; matrix assisted laser desorption ionization; microcalorimetry; nuclear magnetic resonance spectroscopy; protein structure; proteolysis; stable isotope; structural biology; thermodynamics; transcription factor

DESCRIPTION: The goal of this project is an atomic-level description of the mechanism by which a yeast zinc finger transcription factor, ADR1, controls transcription of the ADH2 gene in Saccharomyces cerevisae. Structural, dynamic, and energetic aspects of two processes central to transcriptional activation will be investigated using high resolution NMR spectroscopy, complemented by thermodynamic measurements using microcalorimetry: (1) sequence-specific binding by the DNA binding domain of ADR1 to its cognate DNA, the upstream activator sequence, UAS1, and (2) structural properties of an identified activation domain in ADR1. The initial aims include a complete characterization of the minimal DNA-binding domain of ADR1, in both its free form and when bound to a UAS1 DNA site. The domain structure of the protein will be characterized by limited proteolysis and MALDI-mass spectrometry.

描述：该项目的目标是对 酵母锌指转录因子ADR1的调控机制 ADH2基因在酿酒酵母中的转录结构， 转录中心的两个过程的动态和能量方面 激活将使用高分辨率核磁共振波谱进行研究， 辅以微量热法进行热力学测量：(1) ADR1的DNA结合域与其同源物的序列特异性结合 DNA，上游激活序列，UAS1，和(2)结构性质 ADR1中已识别的激活域。最初的目标包括 ADR1最小DNA结合域的完整表征，在这两个 它的自由形式，当它与UAS1 DNA站点结合时。的域结构 该蛋白质的特征是有限的蛋白质降解和MALDI质量 光谱分析。