The role of Orc6 in DNA replication and cytokinesis.

Orc6 在 DNA 复制和胞质分裂中的作用。

• 批准号: 10581262
• 负责人: IGOR N CHESNOKOV
• 金额: $ 3.62万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581262
• 关键词: Administrative Supplement; Biological Models; Cells; Complex; Cytokinesis; Cytoskeleton; DNA biosynthesis; DNA-Protein Interaction; Detection; Documentation; Drosophila genus; Ensure; Equipment; Eukaryota; Eukaryotic Cell; Fluorescence; Funding; Future; Gel; Goals; Grant; Human; Image; Investments; Performance; Polymers; Positioning Attribute; Postdoctoral Fellow; Protein Complex Subunit; Proteins; Research; Role; Stains; System; Tissues; United States National Institutes of Health; Visible Radiation; Visualization; imager; mutant; parent grant; preservation; protein complex; protein function

This is an application for the Administrative Supplement to the currently active R01 GM121449 grant entitled “The Role of Orc6 in DNA replication and cytokinesis”. The major thrust of the parent grant is to understand the functions of replication initiators Orc6 and ORC in eukaryotic cells and tissues using Drosophila and human cells as model systems. ORC, heteromeric six-subunit protein complex, is essential for DNA replication. The smallest of ORC subunits, Orc6 protein, is important for DNA replication in all eukaryotes. We have shown that in Drosophila Orc6 has an additional role in cytokinesis through interaction with the septin complex, a highly conserved polymerizing protein assembly that is critical for cytokinesis in many species and is recognized as important component of the cytoskeleton. In preliminary studies we have identified structural features of Orc6 that contribute directly to the functions of the protein in replication and cytokinesis. The overall goal of this proposal is to define the distinct mechanisms of Orc6 function in DNA replication and in cytokinesis using Drosophila as a model system. All major objectives of the parent grant require constant purifications of the wild type and mutant protein complexes (such as ORC and septin complex and their subunits) to study protein functions and protein/DNA interactions. ChemiDoc Imagers offer best-in-class performance with ease of use for visible light (RGB) and far red/near infrared (FR/NIR) fluorescence and chemiluminescence detection and all general gel documentation applications. Stain-free imaging enables immediate visualization of proteins without gel staining and instant verification of protein transfer to blots. The Administrative Supplement will allow us to purchase a new ChemiDoc MP that will greatly facilitate and advance our studies and workflow. The purchase of new equipment required for the project and associated research expenses will provide the necessary investment into the research-oriented economy. We believe that the discoveries made with the help of new ChemiDoc MP system funded by the Administrative Supplement will significantly strengthen our chances for obtaining future NIH support through competitive renewal application. This in turn will ensure preservation of the newly created and current research positions supported by the parent grant.

这是对当前有效的 R01 GM121449 补助金的行政补充申请，题为 “Orc6 在 DNA 复制和胞质分裂中的作用”。母基金的主要目的是了解 使用果蝇和人类细胞研究复制起始子 Orc6 和 ORC 在真核细胞和组织中的功能 作为模型系统，ORC，异聚六亚基蛋白质复合物，是 DNA 复制所必需的。 ORC 亚基 Orc6 蛋白对所有真核生物中的 DNA 复制都很重要。 果蝇 Orc6 通过与 septin 复合物相互作用，在胞质分裂中发挥额外作用，septin 复合物是一种高度 保守的聚合蛋白组装，对许多物种的胞质分裂至关重要，被认为是 在初步研究中，我们已经确定了 Orc6 的结构特征。 直接促进蛋白质复制和胞质分裂的功能。 提议是定义 Orc6 在 DNA 复制和胞质分裂中功能的不同机制 果蝇作为模型系统的所有主要目标都需要对野生环境进行不断的纯化。 类型和突变蛋白复合物（例如 ORC 和 septin 复合物及其亚基）来研究蛋白质 ChemiDoc 成像仪提供一流的性能且易于使用。 可见光（RGB）和远红/近红外（FR/NIR）荧光和化学发光检测等 一般凝胶记录应用程序无需染色即可立即可视化蛋白质。 凝胶染色和蛋白质转移到印迹的即时验证将使我们能够 购买新的 ChemiDoc MP，这将极大地促进和推进我们的研究和工作流程。 该项目所需的新设备和相关研究费用将提供必要的 我们相信，新的发现有助于研究型经济。 由行政补充文件资助的 ChemiDoc MP 系统将显着增强我们的机会 通过竞争性更新申请获得未来的 NIH 支持，这反过来将确保保存。 新设立的和现有的研究职位由母公司拨款支持。