Mechanisms for recruitment and function of metazoan replication initiation factors

后生动物复制起始因子的招募和功能机制

• 批准号: 10594040
• 负责人: Franziska Bleichert
• 金额: $ 35.18万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594040
• 关键词: Address; Basal Cell Nevus Syndrome; Binding; Binding Proteins; Biochemical; Biological Assay; Cell Proliferation; Cell Survival; Cells; Chromatin; Chromosomal Duplication; Complex; Cryoelectron Microscopy; DNA; DNA Binding; DNA Sequence; DNA biosynthesis; Data; Dedications; Deposition; Development; Disease; Dwarfism; Environment; Eukaryota; Event; Failure; Genomic Instability; Genomics; Goals; Growth; In Vitro; Knowledge; Label; Laboratories; Learning; Licensing; Life; Link; Maintenance; Malignant Neoplasms; Mediating; Modeling; Molecular; Motor; Mutation; Nucleosomes; Organism; Outcome; Peptide Initiation Factors; Positioning Attribute; Process; Proteins; Regulation; Replication Initiation; Replication Origin; Research; Research Proposals; Saccharomyces cerevisiae; Saccharomycetales; Site; Specific qualifier value; Structure; System; Techniques; Testing; Work; Yeast Model System; biochemical model; cell growth; cofactor; genetic information; helicase; human disease; in vivo; novel; novel therapeutic intervention; origin recognition complex; recruit

Project Summary The survival of living organisms depends on the timely and accurate duplication of chromosomal DNA. In all domains of life, the onset of DNA replication (or initiation) relies on dedicated initiator proteins that bind genomic sites, termed replication origins, and help load replicative helicases onto DNA. In eukaryotes, the initiator is the origin recognition complex (ORC), which recruits and deposits the Mcm2-7 helicase motor module onto DNA to ‘license’ origins. Although core replication initiation factors are conserved across most eukaryotes and are well-studied in budding yeast, important differences exist with respect to origin recognition and the regulation of origin licensing between S. cerevisiae and higher eukaryotes. Numerous outstanding questions therefore remain in the metazoan system regarding origin recruitment and function of core and accessory DNA replication initiation factors, and the specific contributions of chromatin-associated proteins to these events. In this proposal, we aim to define at a mechanistic level in metazoan systems how ORC- dependent DNA remodeling contributes to Mcm2-7 loading, how disease-linked mutations alter initiator activities, and how ORC-partner proteins promote chromosomal recruitment and function of the initiator, by integrating biochemical, structural, and cell-based approaches. The findings from this work will have broad implications for multiple scientific fields, as they will not only help generate models for origin specification, origin processing, and origin licensing in metazoans, but also contribute to our understanding of DNA- and chromatin-dependent macromolecular machines. Our studies are also of significant biomedical relevance since the failure to precisely replicate chromosomal DNA causes genome instability, which in turn underpins many human diseases, including cancer. In the long-term, the outcomes of our efforts thus have the potential to provide important starting points for the development of novel therapeutic strategies.

项目摘要 活生物体的生存取决于染色体DNA的及时复制。总的来说 生命领域，DNA复制（或启动）的发作依赖于结合的专用引发剂蛋白 基因组位点，称为复制起源，并有助于将复制性解旋酶载到DNA上。在真核生物中 引发剂是原点识别综合体（ORC），它报告并沉积MCM2-7旋转酶电动机 模块到DNA到“许可”起源。尽管核心复制计划因素在大多数 真核生物，在萌芽的酵母中进行了充分研究，在起源识别方面存在重要差异 以及酿酒酵母和更高真核生物之间的起源许可的规定。许多杰出 因此，有关核心的起源招聘和功能以及 辅助DNA复制倡议因素，以及与染色质相关蛋白的特定贡献 这些事件。在此提案中，我们的目标是在后生动物系统中定义机械水平。 依赖性DNA重塑有助于MCM2-7加载，疾病连接突变如何改变引发剂 活动以及Orc-Partner蛋白如何促进引发剂的染色体募集和功能 整合生化，结构和基于细胞的方法。这项工作的发现将有广泛的 对多个科学领域的影响，因为它们不仅有助于生成原始规范的模型，还将 起源处理和后生动物的起源许可，但也有助于我们对DNA和DNA的理解 染色质依赖性大分子机器。我们的研究也具有重要的生物医学相关性 无法精确复制染色体DNA导致基因组不稳定性，而这又是许多基础 包括癌症在内的人类疾病。从长远来看，我们努力的结果有可能 为开发新的治疗策略提供重要的起点。