Structure and Function of Bacterial RecN Protein

细菌 RecN 蛋白的结构和功能

• 批准号: 7692795
• 负责人: SHELLEY LUSETTI
• 金额: $ 14.56万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-24 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7692795
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Bacteria; Bacterial Genome; Bacterial Model; Binding; Biochemical; Biology; C-terminal; Chromosomes; Coiled-Coil Domain; Communication Research; Complex; Conserved Sequence; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Pathway; DNA repair protein; Deinococcus radiodurans; Development; Double Strand Break Repair; Educational workshop; Escherichia coli; Genetic; Genome; Goals; Grant; Homologous Gene; Housing; Hydrolysis; In Vitro; Investigation; Kinetics; Laboratories; Maintenance; Manuscripts; Mentorship; Metabolism; Modeling; Molecular; Mutation; Organism; Orthologous Gene; Physical condensation; Pilot Projects; Play; Positioning Attribute; Property; Protein Family; Proteins; Reaction; Recombinant Proteins; Research; Research Personnel; Research Proposals; Role; Sister Chromatid; Structure; System; Testing; Training; Tumor Suppressor Proteins; Universities; Wisconsin; Work; cohesin; cohesion; ds-DNA; enzyme pathway; in vivo; member; post-doctoral training; protein complex; protein protein interaction; public health relevance; recombinase; repaired; stem

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.

描述（由申请人提供）：本研究计划的总体目标是揭示 RecN 的分子功能，RecN 是一种对修复细菌 DNA 双链断裂 (DSB) 至关重要的蛋白质。细菌 RecN 蛋白与染色体结构维持 (SMC) 家族蛋白具有显着的同源性。真核 SMC 蛋白在染色体浓缩、姐妹染色单体凝聚和重组 DNA 修复等多种 DNA 代谢过程中具有重要的（尽管尚未完全了解）管家和肿瘤抑制作用。尽管大量的遗传证据强调了 RecN 蛋白对细菌基因组维护的重要性，但尚未对其功能进行实质性的生化研究，因此，需要 RecN 的特定反应尚未确定。这种情况反映了几个研究小组在研究大肠杆菌（一种模式细菌生物）RecN 蛋白时遇到的困难，原因是该蛋白过度表达的不溶性。然而，我们已经鉴定出一种在大肠杆菌中表达时可溶的 RecN 同源物，并准备启动试点研究，为细菌 RecN 蛋白在重组 DNA 修复途径中的生化功能建立可检验的假设。这项工作将为理解真核 SMC 蛋白在基因组维护中发挥的分子作用提供有价值的模型。 公共卫生相关性：生化方法为了解 SMC 蛋白在基因组维护和重组 DNA 修复中发挥的分子作用提供了有价值的模型。