Replication Initiation in Bacteria and Eukaryotes

细菌和真核生物中的复制起始

• 批准号: 8247781
• 负责人: David Jeruzalmi
• 金额: $ 26.99万
• 依托单位: CITY COLLEGE OF NEW YORK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247781
• 关键词: Address; Antibiotics; Antineoplastic Agents; Architecture; Area; Bacillus cereus; Bacteria; Bacteriophages; Binding; Binding Sites; Biological Assay; Cell Cycle; Cells; Chromosomes; Collection; Complex; DNA; DNA Binding; DNA Primase; DNA biosynthesis; DNA replication origin; Data; Deposition; Development; DnaB helicase; Drug Design; Elements; Ensure; Escherichia coli; Eukaryota; Foundations; Genome; Goals; Health; Human; Length; Maintenance; Malignant Neoplasms; Measures; Modeling; Molecular Target; Nucleotides; Pathway interactions; Positioning Attribute; Proteins; Recruitment Activity; Regulation; Replication Initiation; Research; Roentgen Rays; Sampling; Science; Shapes; Signal Transduction; Site; Structure; Work; X-Ray Crystallography; base; design; helicase; macromolecular assembly; novel; novel therapeutics; origin recognition complex; protein complex; research study; success

DESCRIPTION (provided by applicant): The goal of this work is a structure-based understanding of how DNA replication is initiated. 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. We study replication initiation complexes in eukaryotes and bacteria. A large body of work has described components of origin complexes and details of how they operate. What is missing, however, in any of these cases, is a three-dimensional view of the initiation machine to guide our understanding of how these components work together. The overall goal of our research is to provide such views. The current proposal incorporates three complementary approaches that focus on several components involved in origin recognition and DNA unwinding. Aim #1 is directed at several bacterial helicaseloader complexes, whose structure and function can be dissected at a sophisticated level and will inform our understanding of eukaryotic complexes. Aim #2 is addressed at obtaining and analyzing the complete ORC ensemble. Aim #3 exploits our recent identification of a unique bacterial MCM complex, which is more tractable, but should still provide a faithful model for its eukaryotic counterparts. PUBLIC HEALTH RELEVANCE Cells prepare for the next round of DNA replication by depositing macromolecular assemblies onto sites on the genome termed origins of DNA replication. Emerging evidence suggests that these DNA replication initiation assemblies represent a rich collection of molecular targets against which to design novel therapeutics. This pathway is not well represented in efforts to discover anti-cancer agents. Lack of useful structural information on relevant molecular targets has stymied efforts at structure-guided drug design. Our structure determination efforts will advance efforts to discover novel anti-cancer agents. Also, our work with bacterial complexes will provide much needed targets for development of novel antibiotics.

描述（由申请人提供）：这项工作的目标是基于结构的理解DNA复制是如何启动的。复制开始于基因组上的多个位点，称为DNA复制的起点。专门的蛋白质复合物结合在这些位点上，为复制做准备。这些基因组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、调控和作用机制尚不完全清楚。我们的工作具有实际意义，因为起源复合体的调控变化有助于人类癌症。细菌复合体的研究将为新型抗生素的开发提供急需的靶点。我们研究真核生物和细菌的复制起始复合物。大量的工作已经描述了起源复合体的组成部分以及它们如何运作的细节。然而，在这些情况下，缺少的是启动机器的三维视图，以指导我们理解这些组件如何协同工作。我们研究的总体目标就是提供这样的观点。目前的建议结合了三种互补的方法，重点关注涉及起源识别和DNA解绕的几个组成部分。目的1是针对几种细菌螺旋加载复合物，其结构和功能可以在复杂的水平上解剖，并将告知我们对真核复合物的理解。目标#2旨在获取和分析完整的ORC集成。Aim #3利用我们最近鉴定的一种独特的细菌MCM复合体，这种复合体更容易处理，但仍然应该为真核生物的对应物提供一个忠实的模型。细胞通过将大分子组装体沉积在基因组上称为DNA复制起点的位点上，为下一轮DNA复制做准备。新出现的证据表明，这些DNA复制起始组装代表了一个丰富的分子靶标集合，可以设计新的治疗方法。在发现抗癌药物的努力中，这一途径并没有得到很好的体现。缺乏相关分子靶点的有用结构信息阻碍了结构导向药物设计的努力。我们的结构测定工作将推动发现新型抗癌药物的努力。此外，我们对细菌复合体的研究将为开发新型抗生素提供急需的靶点。

项目成果

A biochemically active MCM-like helicase in Bacillus cereus.

蜡样芽孢杆菌中具有生化活性的 MCM 样解旋酶。

• DOI: 10.1093/nar/gkp376
• 发表时间: 2009
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Samuels,Martin;Gulati,Gaurav;Shin,Jae-Ho;Opara,Rejoice;McSweeney,Elizabeth;Sekedat,Matt;Long,Stephen;Kelman,Zvi;Jeruzalmi,David
• 通讯作者: Jeruzalmi,David