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NMR STRUCTURAL AND BIOPHYSICAL STUDIES OF XRCC1

XRCC1 的 NMR 结构和生物物理研究

基本信息

批准号：
6635490
负责人：
MICHAEL R GRYK
金额：
$ 20.9万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-05-01 至 2006-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6635490
关键词：
DNA damage DNA repair biophysics intermolecular interaction nuclear magnetic resonance spectroscopy protein structure function

项目摘要

DESCRIPTION: (adapted from the applicant's abstract) Xray cross complementing protein 1 (XRCC1) participates in the repair of DNA damage and, in mice, has been indicated to also be required for embryonic development. XRCC1 interacts with the DNA repair proteins poly-ADP-ribose polymerase (PARP), DNA polymerase (beta-Pol), and DNA ligase III. The complex formed between XRCC1, beta-Pol and DNA ligase III repairs incised abasic sites in the final three steps of the base excision repair pathway. The N-terminal domain of XRCC1 has been shown to bind specifically to single-strand break DNA and its structure has recently been determined by high resolution NMR methods. This proposal seeks to characterize the molecular mechanism of the interaction of the N-terminal domain of XRCC1 with DNA substrate and with beta-Pol. Mutagenesis methods, in combination with biochemical assays, will be used to elucidate those residues that contribute to binding. The existing NMR-based structure and chemical shift perturbation map of the binding surface will guide the mutagenesis. The full range of DNA substrate specificity will also be explored. Backbone dynamics will be characterized and correlated with functional binding sites. The 40 kDa complex, involving the N-terminal domain of XRCC1, DNA substrate and beta-Pol, will be structurally characterized using NMR-based methods.

描述：（改编自申请人的摘要）X射线交叉互补蛋白1（XRCC 1）参与DNA损伤的修复，在小鼠中，也被认为是胚胎发育所必需的。XRCC 1相互作用与DNA修复蛋白聚ADP-核糖聚合酶（PARP）、DNA聚合酶（β-Pol）和DNA连接酶III。XRCC 1、β-Pol和 DNA连接酶III修复切割的脱碱基位点在最后三个步骤中，碱基切除修复途径XRCC 1的N-末端结构域已被证明是特异性结合单链断裂DNA，其结构最近通过高分辨率NMR方法测定。这项建议旨在表征N-末端相互作用的分子机制用DNA底物和用β-Pol诱变方法，结合生化分析，将用于阐明这些残留物这有助于约束力。现有的基于NMR的结构和化学位移结合表面的微扰图将指导诱变。充分还将探索DNA底物特异性的范围。主干动力学将被表征并与功能结合位点相关联。40 kDa 复合物，涉及XRCC 1的N-末端结构域、DNA底物和β-Pol，将使用基于NMR的方法进行结构表征。