Regulation of genomic stability by ORC

ORC 调节基因组稳定性

• 批准号: 9753284
• 负责人: Supriya G Prasanth
• 金额: $ 30.09万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753284
• 关键词: Address; Affect; Binding; Binding Proteins; Biochemical; Biological; Biomedical Research; Breast Cancer Patient; Cell Cycle Progression; Cell Proliferation; Cell Survival; Cells; Chromatin; Closure by clamp; Complex; Cytokinesis; DNA; DNA Binding; DNA Damage; DNA biosynthesis; DNA replication fork; Data; Generations; Genome; Genome Stability; Genotoxic Stress; Goals; Heterochromatin; Human; Image; Individual; Kinetochores; Laboratories; Maintenance; Malignant Neoplasms; Mission; Mitosis; Molecular; Oxidative Stress; Pathway interactions; Phosphorylation; Play; Positioning Attribute; Post-Translational Protein Processing; Prognostic Marker; Proteins; Proteome; Public Health; Regulation; Replication Initiation; Replication Licensing; Replication Origin; Research; Resolution; Role; S Phase; System; Techniques; Testing; Threonine; Time; Treatment Efficacy; United States National Institutes of Health; base; biophysical techniques; cancer biomarkers; cell growth; combat; experience; genome integrity; innovation; insight; malignant breast neoplasm; mimetics; mutant; novel; origin recognition complex; oxidative DNA damage; oxidative damage; prevent; repaired; response; scaffold; sensor; single molecule; therapeutic target; tumorigenesis

Maintenance of genomic integrity is paramount to cell survival. Precise duplication of our genome and appropriate cellular response to genotoxic stress is critical to maintain genome stability. Origin Recognition Complex (ORC, composed of six subunits) and ORC-Associated (ORCA) are required for the initiation of DNA replication and regulate heterochromatin organization. Multiple subcomplexes of ORC and/or individual ORC subunits regulate different aspects of cell cycle progression and thus play pivotal roles in the maintenance of genomic stability. The long-term goal of my laboratory is to understand how ORC executes and coordinates various aspects of cell growth, proliferation and survival. The smallest subunit of ORC, Orc6, is required for DNA replication and also coordinates cytokinesis. The scientific premise of this proposal that human Orc6 is required for DNA replication progression and DNA Damage Response is based on strong preliminary data. We have exciting preliminary data demonstrating yet another novel role of Orc6, in DDR. We observe that the levels of Orc6 increase upon DNA damage and that Orc6 undergoes specific phosphorylation during oxidative DNA damage. Further, cells lacking Orc6 fail to activate ATR upon DNA damage. The objective of the present proposal is to answer fundamental questions on the roles of Orc6 in regulating S-phase and during DDR. Our hypothesis, based on preliminary data, is that Orc6 interacts with replication fork components and facilitates DNA replication and upon DNA damage the loss of this interaction inhibits replication fork progression. With our expertise and experience in cell biological and biochemical characterization of Orc6 and strong preliminary data, we are ideally positioned to pursue the following specific aims: 1) Determine the role of Orc6 in replication fork progression. 2) Determine the role of Orc6 in DNA damage response. 3) Determine the role of post-translational modifications on Orc6 in DDR and cell cycle progression. An important question in the field that has remained to be answered is why do ORC proteins remain bound to chromatin in postG1 cells? This proposal is significant because we address this question by studying how Orc6 regulates replication fork progression and it evaluates for the first time the role of preRC proteins in DNA damage response. This proposal is conceptually innovative because we will rigorously dissect novel regulatory mechanisms of Orc6 in DNA replication progression and DNA damage response. This proposal is technologically innovative because it employs state of the art cell biological techniques, including super-resolution imaging combined with biochemical and single molecule biophysical approaches. Understanding how Orc6 governs multiple pathways including DNA replication, mitosis and DDR is expected to uncover novel pathways that would be useful to prevent tumorigenesis and key to allow more effective therapeutic targeting to combat cancer.

保持基因组的完整性对细胞的生存至关重要。我们基因组的精确复制和 适当的细胞对基因毒性应激的反应是维持基因组稳定的关键。起源 需要识别复合体(ORC，由六个亚基组成)和ORC关联(ORCA) DNA复制的启动和异染色质组织的调节。的多个亚复合体 ORC和/或单个ORC亚基调节细胞周期进程的不同方面，从而发挥作用 在维持基因组稳定性方面的关键作用。我的实验室的长期目标是 了解ORC如何执行和协调细胞生长、增殖和 生死存亡。ORC的最小亚基ORC6是DNA复制所必需的，也是坐标 胞质分裂。这一建议的科学前提是DNA复制需要人类ORC6 进展和DNA损伤反应是基于强大的初步数据。我们有令人兴奋的 初步数据表明，ORC6在DDR中还有另一个新的作用。我们观察到， ORC6在DNA损伤时增加，并在氧化过程中经历特异的磷酸化 DNA损伤。此外，缺乏ORC6的细胞在DNA损伤时无法激活ATR。该计划的目标是 目前的建议是回答关于ORC6在调节S阶段和 在DDR期间。基于初步数据，我们的假设是ORC6与复制叉相互作用 成分并促进DNA复制，一旦DNA损伤，这种相互作用的丧失就会抑制 复制分叉进程。凭借我们在细胞生物学和生化方面的专业知识和经验 ORC6的特征和强劲的初步数据，我们处于追求以下目标的理想位置 具体目标：1)确定ORC6在复制分叉进展中的作用。2)确定ORC6的作用 在DNA损伤反应中。3)确定ORC6的翻译后修改在DDR和 细胞周期进程。该领域一个有待回答的重要问题是，为什么要 ORC蛋白在后G1细胞中仍与染色质结合？这项建议意义重大，因为我们 通过研究ORC6如何调节复制分叉进程以及它对 首次研究了前RC蛋白在DNA损伤反应中的作用。这项建议在概念上是 创新是因为我们将严格剖析ORC6在DNA复制中的新调控机制 进展和DNA损伤反应。这项提议在技术上是创新的，因为它采用了 最先进的细胞生物技术，包括超分辨率成像与生化相结合 和单分子生物物理方法。了解ORC6如何控制多条途径 包括DNA复制、有丝分裂和DDR有望发现有用的新途径 预防肿瘤发生和使更有效的治疗靶向抗击癌症的关键。