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高度保守。广泛的研究支持了与IA型拓扑异构酶一致的RECQ解旋酶参与基因组稳定性的维持。但是，其分子基础尚未清楚地确​​定。与许多单个细胞生物不同，例如大肠杆菌，酿酒酵母和糖疗中的葡萄糖，它们具有一种或两种IA型拓扑异构酶和单一RECQ解旋酶，某些革兰氏阳性细菌，例如cereus cereus andthracis of Etharacis of类chromax，chromagial of Ethracis of Typeries of Typeries of The类药物。拓扑异构酶（在蜡状芽孢杆菌中被注释为Bctopo I，Bctopo III？和Bctopo III？）和RecQ解旋酶的两个染色体拷贝（在Cereus bacillus bacillus sanceps of BcRecQs和BcRecql注释）。在蜡状芽孢杆菌中，Bctopo III之间的物理相互作用？已经确定了BCRECQL。 Bctopo III的表征？使用高度纯化的活性酶和可行的遗传系统和BCRECQL蛋白复合物将揭示基因组稳定性IA型IA型IA型topoasomerase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 DNA代谢过程，以及其他细菌特异性DNA代谢过程，例如转播和posposonativation，plobsentional contresention，plosposcrect，plosposcrect和sej conj and sej conj and sej conj and sej。此外，从这些研究中获得的见解可能会阐明细菌如何应对诸如抗生素，化学物质，紫外线和自由基等严重的环境压力。已经提出了以下特定目标来阐明BCTOPO III的功能？和BCRECQL蛋白复合物。具体目的是：1）隔离Bctopo III？和BCRECQL蛋白复合物，并表征其生化特性。 2）映射和表征Bctopo III的域？和BCRECQL对于相互作用至关重要。 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会揭示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。这些研究还可能使靶向细菌IA型拓扑异构酶和RECQ解旋酶有益于抗生素设计。