MECHANISMS OF HMG-BOX-DNA RECOGNITION

HMG-BOX-DNA 识别机制

• 批准号: 6019285
• 负责人: Rachel E Klevit
• 金额: $ 18.32万
• 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: 1997-09-30 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6019285
• 关键词: DNA binding protein; DNA footprinting; animal genetic material tag; electron spin resonance spectroscopy; intermolecular interaction; nuclear magnetic resonance spectroscopy; nucleic acid sequence; thermodynamics; transcription factor

DESCRIPTION: This is a new proposal aimed at obtaining information on the structural, thermodynamic, and dynamic aspects of non-sequence specific modes of protein-nucleic acid recognition by the HMG-box motif. The objective will be to understand the recognition in terms of two limiting mechanisms: recognition of preformed structure (lock and key) vs induction of structure in inherently flexible DNA sequences (induced fit). A multifaceted approach is proposed in order to give the most complete picture: electrophoretic mobility shift assays and radical footprinting for characterizing binding of different DNA sequences, isothermal titration calorimetry for determining thermodynamic parameters, EPR for determining DNA dynamics, and NMR for structure analysis of the proteins and its complexes with DNA. The goal is to characterize the binding of nucleic acids containing one each of ~7 structural and/or dynamical features to a single HMG-box from the Xenopus Laevis Upstream Binding Factor (xUBFBox1).

说明：这是一项新的提案，旨在获得关于 非序列特异性的结构、热力学和动力学方面 HMG-盒基序的蛋白质-核酸识别模式。 的 目标将是理解两个限制方面的承认 机制：识别预成型结构（锁和钥匙）与感应 在固有的灵活的DNA序列的结构（诱导适合）。 一 提出了多方面的方法，以提供最完整的 图片：电泳迁移率变动分析和自由基足迹 表征不同DNA序列的结合，等温滴定 量热法测定热力学参数，EPR测定 DNA动力学和NMR用于蛋白质及其 与DNA的复合物。 目标是表征核酸的结合， 酸含有~7种结构和/或动力学特征中的每一种， 来自非洲爪蟾上游结合因子（xUBFBox 1）的单个HMG盒。