Regulation of Membrane Phospholipid Synthesis

膜磷脂合成的调控

• 批准号: 6987191
• 负责人: ANIBAL SOTO-CARDALDA
• 金额: $ 3.61万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6987191
• 关键词: SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; enzyme activity; fungal genetics; gel mobility shift assay; genetic promoter element; membrane reconstitution /synthesis; messenger RNA; northern blottings; phospholipids; phosphorylation; polymerase chain reaction; predoctoral investigator; protein kinase; reporter genes; scintillation counter; transcription factor; western blottings; zinc

DESCRIPTION (provided by applicant):The yeast Saccharomyces cerevisiae serves as a model eukaryote to study the regulation of membrane phospholipid synthesis. The genes and enzymes in the phospholipid synthesis pathways are regulated by genetic (e.g., zinc depletion) and biochemical (e.g., phosphorylation) mechanisms. Recent studies have shown that phospholipid synthesis is regulated by the stress condition of zinc depletion. Zinc is an essential element required for the optimal growth and metabolism of eukaryotic cells. Preliminary data indicated that a pathway for synthesizing phosphatidylcholine, the most abundant membrane phospholipid, was activated to allow cells to deal with this stress. In this application, I will examine the regulation of CK/7-encoded choline kinase, the enzyme that catalyzes the committed in the Kennedy pathway, in response to zinc depletion. I will measure the levels of choline kinase activity, protein, and mRNA. Transcription of the CKI1 gene will be monitored using a Pc/c/r/acZ reporter gene. Identification of the promoter element(s) and transcription factor(s) involved will be pursued. The phosphorylation state of choline kinase in response to zinc depletion will be monitored by in vivo labeling of the protein with 32P, using wild type and available phosphorylation site mutants.

描述（由申请人提供）：酵母酿酒酵母作为模型真核生物，用于研究膜磷脂合成的调节。磷脂合成途径中的基因和酶受遗传（例如，锌耗尽）和生化（例如，磷酸化）机制。最近的研究表明，磷脂的合成受到锌缺乏的应激条件的调节。锌是真核细胞最佳生长和代谢所需的必需元素。初步数据表明，合成磷脂酰胆碱（最丰富的膜磷脂）的途径被激活，使细胞能够应对这种压力。在本申请中，我将研究CK/7编码的胆碱激酶的调节，该酶催化肯尼迪途径中的承诺，以响应锌的耗尽。我将测量胆碱激酶活性，蛋白质和mRNA的水平。将使用Pc/c/r/acZ报告基因监测CKI 1基因的转录。将对所涉及的启动子元件和转录因子进行鉴定。胆碱激酶对锌耗竭的磷酸化状态将通过使用野生型和可用的磷酸化位点突变体用32 P体内标记蛋白质来监测。