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.

这是当前活跃的R01 GM121449赠款的管理补充的申请 “ ORC6在DNA复制和细胞因子中的作用”。父母赠款的主要目的是了解 使用果蝇和人类细胞在真核细胞和组织中复制启动器ORC6和ORC的功能 作为模型系统。兽人，杂物六亚基蛋白复合物，对于DNA复制至关重要。最小 ORC亚基的ORC6蛋白对于所有真核生物中的DNA复制都很重要。我们已经证明了这一点 果蝇ORC6通过与Septin复合物相互作用在细胞因子中具有附加作用 保守的聚合蛋白质组件，对于许多物种中的细胞因子至关重要，被认为是 细胞骨架的重要组成部分。在初步研究中，我们已经确定了ORC6的结构特征 这直接有助于蛋白质在复制和细胞因子中的功能。总体目标 建议是在使用DNA复制和细胞因子中定义ORC6功能的不同机制 果蝇作为模型系统。父母赠款的所有主要对象都需要持续净化野外 类型和突变蛋白复合物（例如ORC和Septin复合物及其亚基）研究蛋白 功能和蛋白质/DNA相互作用。 Chemidoc图像可轻松使用一流的性能 可见光（RGB）和远红色/近红外（FR/NIR）荧光和化学发光检测以及所有 一般凝胶文档应用。无染色成像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。行政补充将使我们能够 购买新的Chemidoc MP，将极大地支持并推进我们的研究和工作流程。购买 项目和相关研究费用所需的新设备将提供必要的 投资以研究为导向的经济。我们认为，在新的帮助下发现的发现 由行政补充资助的Chemidoc MP系统将大大增强我们的机会 通过竞争性更新应用获得未来的NIH支持。反过来将确保保存 由父母赠款支持的新创建和当前的研究职位。