X-RAY ANALYSIS OF REPLICATION AND REPAIR PROTEINS

复制和修复蛋白的 X 射线分析

• 批准号: 8361624
• 负责人: David Jeruzalmi
• 金额: $ 4.02万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361624
• 关键词: Antibiotics; Architecture; Binding; Cell Cycle; Complex; DNA; DNA Sequence; DNA replication origin; Development; Dissociation; Ensure; Funding; Genome; Goals; Grant; Human; Lead; Left; Lesion; Malignant Neoplasms; Monitor; National Center for Research Resources; Nucleotide Excision Repair; Principal Investigator; Process; Proteins; Reaction; Recruitment Activity; Regulation; Replication Initiation; Research; Research Infrastructure; Resources; Roentgen Rays; Side; Site; Source; Structure; Surgical incisions; United States National Institutes of Health; Work; base; cost; endonuclease; novel; protein complex; repaired; restoration; structural biology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The goal of this work is a structure-based understanding of 1) replication initiation and 2) nucleotide excision repair (NER). Replication initiates at multiple sites on the genome called origins of DNA replication. Specialized protein complexes bind at these sites and prepare the duplex for replication. The activity of these ensembles is tightly controlled to ensure that only one copy of the genome is made per cell cycle. The architecture, regulation and mechanisms of action of these large complexes are incompletely understood. Our work is of practical significance because regulatory changes in origin complexes contribute to human cancers. Efforts with bacterial complexes will provide much needed targets for development of novel antibiotics. The first steps in NER are performed by three proteins: UvrA, UvrB and UvrC. The UvrA+UvrB ensemble monitors DNA and recognizes damage. On encountering damage, UvrA exits the complex, leaving UvrB stably bound at or near the lesion site. Damage searching, formation of the DNA complex and dissociation of UvrA are regulated by ATP. UvrB then recruits the endonuclease UvrC, which catalyzes incisions on either side of the lesion. Additional processing reactions lead to restoration of the original DNA sequence.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 这项工作的目标是基于结构的理解1）复制起始和2）核苷酸切除修复（NER）。复制起始于基因组上的多个位点，称为DNA复制起点。专门的蛋白质复合物在这些位点结合，并准备复制的双链体。这些集合的活性受到严格控制，以确保每个细胞周期只产生一个基因组拷贝。这些大型复合物的结构、调节和作用机制尚不完全清楚。我们的工作具有实际意义，因为起源复合物的调节变化有助于人类癌症。对细菌复合物的研究将为开发新型抗生素提供急需的靶点。 NER中的第一步由三种蛋白质执行：UvrA，UvrB和UvrC。UvrA+UvrB集合监测DNA并识别损伤。在遇到损伤时，UvrA退出复合物，留下UvrB稳定地结合在损伤部位处或附近。损伤搜索、DNA复合物的形成和UvrA的解离受ATP调节。然后UvrB募集核酸内切酶UvrC，其催化损伤两侧的切口。额外的加工反应导致原始DNA序列的恢复。