STRUCTURAL STUDIES OF TRANSCRIPTIONAL ACTIVATION

描述：本项目的目标是对 酵母锌指转录因子ADR 1控制 在酿酒酵母中ADH 2基因的转录。 结构上， 动态和能量方面的两个过程的核心转录 将使用高分辨率NMR光谱学研究活化， 通过使用微量热法的热力学测量补充：（1） ADR 1的DNA结合结构域与其同源物的序列特异性结合 DNA，上游激活序列，UAS 1，和（2）的结构特性 ADR 1中的一个识别的激活域。 最初的目标包括： ADR 1的最小DNA结合结构域的完整表征， 它的游离形式和当结合到UAS 1 DNA位点时。 的结构域结构 蛋白质的特征在于有限的蛋白水解和MALDI-质量 光谱法

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.