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.

项目概要 生物体的生存依赖于染色体DNA的及时、准确的复制。 在生命领域，DNA 复制（或起始）的开始依赖于结合的专用起始蛋白 基因组位点，称为复制起点，有助于将复制解旋酶加载到 DNA 上。 引发剂是起源识别复合物 (ORC)，它招募并沉积 Mcm2-7 解旋酶马达 尽管核心复制起始因子在大多数情况下都是保守的。 真核生物，并在芽殖酵母中进行了充分研究，但在起源识别方面存在重要差异 以及酿酒酵母和高等真核生物之间的起源许可监管。 因此，关于后生动物系统的核心和功能的起源招募和功能仍然存在问题。 辅助 DNA 复制起始因子，以及染色质相关蛋白对复制的具体贡献 在本提案中，我们的目标是在后生动物系统的机械层面上定义 ORC- 依赖性 DNA 重塑有助于 Mcm2-7 加载，疾病相关突变如何改变启动子 活性，以及​​ ORC 伴侣蛋白如何促进染色体募集和启动子功能， 这项工作的结果将具有广泛的整合生化、结构和细胞的方法。 对多个科学领域的影响，因为它们不仅有助于生成原产地规范模型， 后生动物的原产地加工和原产地许可，也有助于我们对 DNA 和 我们的研究也具有重要的生物医学相关性，因为 无法精确复制染色体 DNA 导致基因组不稳定，这反过来又支撑了许多 因此，从长远来看，我们的努力成果有可能改善人类疾病，包括癌症。 为开发新的治疗策略提供重要的起点。