Structure and Function of Bacterial RecN Protein

描述（由申请人提供）：本研究提案的总体目标是揭示RecN的分子功能，RecN是一种对细菌中DNA双链断裂（DSB）修复至关重要的蛋白质。细菌RecN蛋白与染色体结构维持（SMC）蛋白家族具有显著的同源性。真核SMC蛋白在多种DNA代谢过程中具有重要的（尽管尚未完全理解）管家和肿瘤抑制作用，如染色体凝聚，姐妹染色单体凝聚和重组DNA修复。虽然广泛的遗传证据强调了RecN蛋白对细菌基因组维护的重要性，但对其功能尚未进行实质性的生化研究，因此尚未确定需要RecN的特定反应。这种情况反映了几个研究小组在使用大肠杆菌（一种模式细菌生物）RecN蛋白时遇到的困难，这是由于过表达蛋白的不溶性。然而，我们已经鉴定了RecN同源物，其在E.大肠杆菌，并定位于启动试点研究，以建立可测试的假设，细菌RecN蛋白的生化功能，在重组DNA修复途径。这项工作将提供有价值的模型，了解真核SMC蛋白在基因组维护中发挥的分子作用。 公共卫生相关性：生物化学方法为理解SMC蛋白在基因组维护和重组DNA修复中的分子作用提供了有价值的模型。

DESCRIPTION (provided by applicant): The overall goal of this research proposal is to reveal the molecular function of RecN, a protein critical to the repair of DNA double-stranded breaks (DSBs) in bacteria. Bacterial RecN proteins share significant homology to the Structural Maintenance of Chromosomes (SMC) family of proteins. Eukaryotic SMC proteins have essential (although not fully understood) house keeping and tumor suppressor roles in a variety of DNA metabolic processes such as chromosomal condensation, sister chromatid cohesion and recombinational DNA repair. Although extensive genetic evidence underscores the importance of RecN proteins to bacterial genome maintenance, there has been no substantive biochemical investigation into their function and, consequently, the specific reactions requiring RecN have not been identified. This situation reflects the difficulties encountered in working with the Escherichia coli (a model bacterial organism) RecN protein by several research groups due to insolubility of the over-expressed protein. However, we have identified a RecN homolog that is soluble when expressed in E. coli and are positioned to initiate pilot studies to establish testable hypotheses for the biochemical function of bacterial RecN proteins in recombinational DNA repair pathways. This work will provide valuable models for understanding the molecular role that eukaryotic SMC proteins play in genome maintenance. PUBLIC HEALTH RELEVANCE: Biochemical approaches to provide valuable models for understanding the molecular role that SMC proteins play in genome maintenance and recombinational DNA repair.