MECHANISMS OF HMG-BOX-DNA RECOGNITION

描述：这是一项新的建议，旨在获取关于 非特定序列的结构、热力学和动力学方面 HMG-box基序识别蛋白质-核酸的模式。这个 目的将从两个方面理解认识上的限制 机制：对预制结构(锁和钥匙)的识别与归纳 在固有的柔性DNA序列中的结构(诱导FIT)。一个 多方面的方法被提出，以便给出最完整的 图片：电泳迁移率变化分析和自由基足迹 等温滴定表征不同DNA序列的结合 测定热力学参数用的量热计，测定用的EPR DNA动力学和核磁共振技术用于蛋白质及其结构分析 与DNA形成复合体。我们的目标是描述原子核的结合 含有~7种结构和/或动力学特征中的一种的酸 非洲爪哇上游结合因子(XUBFBox1)中的单个HMG-box。

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).