RUI: Cellular Role of CTP:Phosphocholine Cytidylyltransferase Isoforms

RUI：CTP 的细胞作用：磷酸胆碱胞苷酰转移酶亚型

• 批准号: 0343534
• 负责人: Jon Friesen
• 金额: --
• 依托单位: Board of Trustees of Illinois State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0343534&HistoricalAwards=false
• 关键词: RUI; Cellular; Role; CTP; Phosphocholine

The objective of this research project is to understand how the enzyme CTP:phosphocholine cytidylyltransferase (CCT) regulates cellular phosphatidylcholine synthesis. CCT is the rate limiting enzyme of the CDP-choline pathway and therefore is extensively regulated. Translocation of soluble CCT to the surface of a cellular membrane results in enzyme activation, a reversible process that can be mimicked in vitro through association of purified CCT with synthetic lipid vesicles. There are two CCT isoforms present in higher eukaryotes, including Drosophila melanogaster and humans. This research project is designed to determine the role(s) of each individual enzyme isoform within a eukaryotic cell. RNA interference using cultured Drosophila melanogaster Schneider 2 cells will be employed to selectively "knock out" each CCT isoform. The first research objective is to identify the cellular location of each Drosophila melanogaster CCT isoform. A second objective is to determine whether each CCT isoform is essential for cell growth. The final research objective involves elucidation of the contribution of each CCT isoform to phosphatidylcholine synthesis. Completion of the research will provide information about the relative contribution of each CCT isoform to cellular phosphatidylcholine synthesis, elucidate additional roles of CCT within a eukaryotic cell, and provide insight into the necessity of multiple CCT isoforms for the function of eukaryotes. Broad Impact: A broader impact of the research is manifested in the positive effect it will have on the research development of undergraduate students and the educational environment of Illinois State University, a predominantly undergraduate institution. Support for undergraduate Biochemistry majors to conduct full-time summer research fosters a scientific career at a critical, early stage. Independent laboratory research is a valuable complement to the extensive classroom training undergraduate students obtain, allowing application of the scientific knowledge learned in the classroom. In addition, students will obtain technical training in the use of specialized, modern scientific equipment. The result will be a graduate who is well-prepared to begin pursuit of a graduate degree in science.

本研究项目的目的是了解酶CTP：磷酸胆碱胞苷酰转移酶（CCT）如何调节细胞磷脂酰胆碱合成。 CCT是CDP-胆碱途径的限速酶，因此受到广泛调节。 可溶性CCT易位至细胞膜表面导致酶活化，这是一种可逆过程，可通过纯化CCT与合成脂质囊泡的结合在体外模拟。 在高等真核生物中存在两种CCT同工型，包括黑腹果蝇和人类。 本研究项目旨在确定真核细胞内每个酶同工型的作用。 使用培养的黑腹果蝇Schneider 2细胞的RNA干扰将用于选择性地“敲除”每个CCT同种型。 第一个研究目标是确定每个果蝇CCT亚型的细胞定位。 第二个目的是确定每种CCT亚型是否对细胞生长至关重要。 最终的研究目标是阐明每种CCT亚型对磷脂酰胆碱合成的贡献。 研究的完成将提供有关每个CCT亚型对细胞磷脂酰胆碱合成的相对贡献的信息，阐明CCT在真核细胞内的其他作用，并深入了解多个CCT亚型对真核生物功能的必要性。 广泛影响：一个更广泛的影响的研究表现在积极的影响，它将有本科生的研究发展和伊利诺伊州州立大学，一个主要的本科院校的教育环境。 支持本科生生物化学专业进行全日制暑期研究，在关键的早期阶段培养科学事业。 独立的实验室研究是对本科生获得的广泛课堂培训的宝贵补充，可以应用在课堂上学到的科学知识。 此外，学生将获得使用专业，现代科学设备的技术培训。 其结果将是一个毕业生谁是充分准备开始追求研究生学位的科学。