Enhancing IDeA Research Partnerships in Oklahoma

加强俄克拉荷马州 IDeA 研究合作伙伴关系

• 批准号: 10399034
• 负责人: DARRIN R. AKINS
• 金额: $ 13.66万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-06 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10399034
• 关键词: Address; Administrative Supplement; Binding; C-terminal; Catalytic Domain; Cell Cycle; Cell Cycle Progression; Cell Line; Cell Survival; Cells; Centers of Research Excellence; Chromosome abnormality; Collaborations; Competence; DNA Damage; DNA Polymerase II; DNA Repair; DNA biosynthesis; DNA replication fork; Defect; Development; Disease; Endogenous Factors; Ensure; Enzymes; Eukaryota; Exhibits; Future; G1 Phase; Generations; Genes; Genome; Genome Stability; Genomic Instability; Goals; Human; Learning; Link; MCM10 gene; Maintenance; Malignant Neoplasms; Molecular; Molecular Biology Techniques; Mutation; Oklahoma; Pathway interactions; Phosphotransferases; Play; Polymerase; Proteins; Recovery; Replication Error; Research; Research Personnel; Role; S Phase; Saccharomyces cerevisiae; Saccharomycetales; Science; Series; Signal Transduction; Stress; Testing; Therapeutic; Universities; Work; Yeasts; anticancer research; base; genome integrity; innovation; insight; mutant; novel; programs; repaired; replication stress; response; sensor; undergraduate education; undergraduate research; undergraduate student

Title: Deciphering the roles of Mcm10 and Polymerase epsilon interaction in maintaining genome stability. Abstract Maintenance of genome stability is key to cell survival. Defects in DNA replication and errors in the DNA damage response pathway contribute to genome instability. Replication stress is a leading cause of genome instability and occurs when replication forks progress slowly or stall. Intricate checkpoint pathways operate to ensure that the entry or progression through the S phase is blocked when the cells encounter DNA damage. Despite the importance of the checkpoint pathways, our understanding of how perturbed forks are sensed by the checkpoint and hence protected by the activated checkpoint remains far from complete. The long-term goal of our research is to address this lack of information by investigating the molecular interactions between the replication machinery and checkpoint pathways that govern checkpoint signaling and fork stabilization. DNA polymerase epsilon (Polε) and Minichromosome maintenance protein 10 (Mcm10) play important roles in replication, but issues related to these roles in replication remain to be resolved. In addition, there is no evidence that Mcm10 is involved in checkpoint activation. Our preliminary studies in budding yeast suggest that Mcm10 interacts with the essential, conserved C-terminal domain of Pol2 (catalytic subunit of Polε) and activates the checkpoint kinase at the perturbed forks. Here, we propose highly innovative ideas and experimental approaches to unambiguously establish the role of Mcm10 and Polε interaction in human cells at the damaged forks. Our first aim will focus on understanding how MCM10 and POLE1 interaction regulates replisome formation and fork progression during normal replication. This will help identify novel molecular mechanisms involved in DNA replication. The second aim will evaluate MCM10 and POLE1 interaction under DNA damaging conditions by studying checkpoint activation, replication competence, chromosomal aberrations, and DNA repair. Together, these proposed aims will resolve the outstanding issues related to the role of Mcm10 and Polε in DNA replication and checkpoint activation and thereby provide a holistic view of how they promote genome stability. The proposed research will involve a close collaboration between an INBRE investigator and a COBRE investigator and the integration of research with undergraduate education. In addition, this project will provide undergraduate students with the opportunity to learn current molecular biology techniques at a research-focused university. Relevance The aims of this proposal are to determine the roles of Mcm10 and Polε in DNA replication and checkpoint activation pathway. These studies will advance our understanding of how genome integrity is maintained over generations and how this integrity can be disrupted under stress in all eukaryotes.

标题：破译 Mcm10 和聚合酶 epsilon 相互作用在维持 基因组稳定性。 抽象的 维持基因组稳定性是 DNA 复制缺陷和错误的关键。 DNA 损伤反应途径导致基因组不稳定。 基因组不稳定的主要原因，当复制叉进展缓慢或停滞时就会发生。 复杂的检查点通路运作以确保进入或进展通过 S 阶段 尽管检查点很重要，但当细胞遇到 DNA 损伤时，它就会被阻断。 路径，我们对检查点如何感知扰动分叉的理解，从而 受激活的检查点保护我们的长期目标还远未完成。 研究的目的是通过研究之间的分子相互作用来解决信息的缺乏。 控制检查点信号和分叉的复制机制和检查点路径 DNA 聚合酶 epsilon (Polε) 和微型染色体维持蛋白 10 的稳定性。 (Mcm10) 在复制中发挥重要作用，但与复制中这些角色相关的问题仍然存在 此外，没有证据表明Mcm10参与了检查点激活。 我们对芽殖酵母的初步研究表明，Mcm10 与必需的、 Pol2 的保守 C 端结构域（Polε 的催化亚基）并激活检查点 在这里，我们提出了高度创新的想法和实验。 明确建立 Mcm10 和 Polε 相互作用在人类细胞中的作用的方法 我们的首要目标是了解 MCM10 和 POLE1 的相互作用。 在正常复制过程中调节复制体的形成和分叉进展，这将有所帮助。 第二个目标是评估 DNA 复制中涉及的新分子机制。 通过研究检查点在 DNA 损伤条件下 MCM10 和 POLE1 相互作用 激活、复制能力、染色体畸变和 DNA 修复。 拟议的目标将解决与 Mcm10 和 Polε 在 DNA 中的作用相关的悬而未决的问题 复制和检查点激活，从而提供它们如何促进的整体视图 基因组稳定性。 拟议的研究将涉及 INBRE 研究者和 COBRE 研究员以及研究与本科教育的整合。 该项目将为本科生提供学习当前分子生物学的机会 以研究为重点的大学的生物学技术。 关联 该提案的目的是确定 Mcm10 和 Polε 在 DNA 复制和 这些研究将增进我们对基因组如何激活的理解。 完整性是代代相传的，而这种完整性如何在压力下被破坏 真核生物。