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相互作用的作用 基因组稳定性。 抽象的 基因组稳定性的维持是细胞存活的关键。 DNA复制和错误的缺陷 DNA损伤反应途径有助于基因组不稳定性。复制应力是 基因组不稳定性的主要原因，并在复制叉缓慢或失速时发生。 错综复杂的检查点途径可确保进入或进展通过S相 当细胞遇到DNA损伤时被阻止。尽管检查站很重要 途径，我们对扰动叉的理解是由检查站感知的，因此 受激活检查点保护仍然远未完成。我们的长期目标 研究是通过研究分子相互作用来解决这种缺乏信息 控制检查点信号和叉子的复制机械和检查点途径 稳定。 DNA聚合酶Epsilon（POLε）和微小染色体维持蛋白10 （MCM10）在复制中扮演重要角色，但与这些复制角色有关的问题仍然存在 要解决。此外，没有证据表明MCM10参与检查点激活。 我们在萌芽酵母上的初步研究表明，MCM10与必需的， POL2的保守C末端结构域（POLε的催化亚基）并激活检查点 激动叉的激酶。在这里，我们提出了高度创新的想法和实验性 明确确定MCM10和POLε相互作用在人类细胞中的作用的方法 受损的叉子。我们的第一个目标将重点在于了解MCM10和POL1的相互作用 调节正常复制过程中的复制体形成和叉进程。这将有所帮助 确定与DNA复制有关的新型分子机制。第二个目标将评估 通过研究检查点，在DNA破坏条件下的MCM10和POL1相互作用 激活，复制能力，染色体畸变和DNA修复。在一起，这些 拟议的目标将解决与MCM10和POLε在DNA中的作用相关的未来问题 复制和检查点激活，从而为它们如何促进提供了整体观点 基因组稳定性。 拟议的研究将涉及近亲调查员与 山毛调查员和研究与本科教育的整合。此外， 该项目将为本科生提供学习当前分子的机会 以研究为重点大学的生物学技术。 关联 该提案的目的是确定MCM10和POLε在DNA复制中的作用 检查点激活途径。这些研究将促进我们对基因组的理解 保持诚信是世代相传的，以及如何在所有人的压力下破坏这种完整性 真核生物。