Interaction between B. cereus RecQ helicase and topoisomerase III

B. cereus RecQ 解旋酶和拓扑异构酶 III 之间的相互作用

• 批准号: 8005161
• 负责人: Zhiyu Li
• 金额: $ 2.5万
• 依托单位: UNIVERSITY OF THE SCIENCES PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-29 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8005161
• 关键词: Address; Anthrax disease; Antibiotic Resistance; Antibiotics; Bacillus anthracis; Bacillus cereus; Bacteria; Bacterial Spores; Bacterial Typing; Biochemical; Biochemical Genetics; Biological; Biological Process; Characteristics; Chemicals; Complex; DNA; DNA Repair; DNA Structure; Endocytosis; Enterococcus; Enzymes; Escherichia coli; Eukaryotic Cell; Exhibits; Fission Yeast; Free Radicals; Genes; Genetic; Genome Stability; Germination; Gram-Positive Bacteria; Human; Human body; Invaded; Light; Maintenance; Maps; Metabolic Pathway; Metabolism; Molecular; Organism; Pattern; Plasmids; Process; Property; Proteins; Reproduction spores; Saccharomyces; Saccharomyces cerevisiae; Staphylococcus epidermidis; Stress; System; Topoisomerase; Topoisomerase III; Work; Yeasts; antibiotic design; base; coping; helicase; in vivo; insight; pathogen; protein complex; repaired; segregation; tool; ultraviolet lesions

DESCRIPTION (provided by applicant): RecQ helicases and type IA topoisomerases are highly conserved from bacteria to humans. Extensive studies have supported that RecQ helicases in concert with type IA topoisomerases are involved in the maintenance of genome stability; however, its molecular basis hasn't been clearly established. Unlike many single cellular organisms, such as Escherichia coli, Saccharomyces cerevisiae, and Saccharomyces pombe, which possess one or two type IA topoisomerases and single RecQ helicase, some Gram-positive bacteria such as Bacillus cereus and Bacillus anthracis, the etiologic agent of anthrax, encode multiple chromosomal copies of type IA topoisomerase (annotated as bcTopo I, bcTopo III? and bcTopo III? in Bacillus cereus) and two chromosomal copies of RecQ helicase (annotated as bcRecQS and bcRecQL in Bacillus cereus). In Bacillus cereus, a physical interaction between bcTopo III? and bcRecQL has been identified. Characterization of the bcTopo III? and bcRecQL protein complex using highly purified active enzymes and a feasible genetic system will reveal the biochemical and biological functions of Bacillus cereus type IA topoisomerase and RecQ helicase in genomic stability, as well as other bacterial specific DNA metabolic processes, such as transposon integration, plasmid maintenance and segregation, and DNA conjugational mobilization. Furthermore, insights gained from these studies may shed light on how bacteria cope with severe environmental stress such as antibiotics, chemicals, UV and free radicals. The following specific aims have been proposed to elucidate the functions of bcTopo III? and bcRecQL protein complex. Specific Aims are: 1) To isolate the bcTopo III? and bcRecQL protein complex and characterize its biochemical properties. 2) To map and characterize domains of bcTopo III? and bcRecQL that are essential for the interaction. 3) To study the interaction between bcTopo III? and bcRecQL in vivo and investigate its biological consequences. Insights gained from this project may shed light on understating how Gram-positive pathogens cope with severe environmental stress and develop antibiotic resistance. These studies may also benefit antibiotic design targeting bacterial type IA topoisomerase and RecQ helicase.

描述（由申请人提供）：RecQ解旋酶和IA型拓扑异构酶从细菌到人类都是高度保守的。大量研究表明 RecQ 解旋酶与 IA 型拓扑异构酶共同参与维持基因组稳定性；然而，其分子基础尚未明确。与许多单细胞生物（如大肠杆菌、酿酒酵母和粟酒酵母）不同，它们拥有一种或两种 IA 型拓扑异构酶和单一 RecQ 解旋酶，一些革兰氏阳性细菌，如蜡样芽孢杆菌和炭疽杆菌（炭疽病原菌），编码 IA 型拓扑异构酶的多个染色体拷贝（注释为蜡样芽孢杆菌中的 bcTopo I、bcTopo III? 和 bcTopo III?）以及 RecQ 解旋酶的两个染色体拷贝（在蜡样芽孢杆菌中注释为 bcRecQS 和 bcRecQL）。在蜡状芽孢杆菌中，bcTopo III 之间存在物理相互作用？并且 bcRecQL 已被识别。 bcTopo III 的特性？使用高度纯化的活性酶和可行的遗传系统的bcRecQL蛋白复合物将揭示蜡样芽孢杆菌IA型拓扑异构酶和RecQ解旋酶在基因组稳定性中的生化和生物学功能，以及其他细菌特异性DNA代谢过程，例如转座子整合、质粒维持和分离，以及 DNA 接合动员。此外，从这些研究中获得的见解可能有助于了解细菌如何应对抗生素、化学品、紫外线和自由基等严重的环境压力。为了阐明 bcTopo III 的功能，提出了以下具体目标？和 bcRecQL 蛋白质复合物。具体目标是： 1) 分离 bcTopo III？和 bcRecQL 蛋白复合物并表征其生化特性。 2) 绘制并表征 bcTopo III 的域？和 bcRecQL 是交互所必需的。 3）研究bcTopo III之间的相互作用？和 bcRecQL 体内并研究其生物学后果。从该项目中获得的见解可能有助于了解革兰氏阳性病原体如何应对严重的环境压力并产生抗生素耐药性。这些研究也可能有利于针对细菌 IA 型拓扑异构酶和 RecQ 解旋酶的抗生素设计。