REGULATION OF PHOSPHOLIPID SYNTHESIS

在酿酒酵母中，CTP是所有膜的前体 磷脂酰乙醇胺甲基化合成磷脂 (主要)和基于CDP-乙醇胺和CDP-胆碱的(辅助) 小路。这项拨款申请解决了细胞 CTP水平通过初级和间接调节磷脂的合成 通过控制蛋白的表达和/或活性来辅助通路 这些途径中的酶。CTP对磷脂合成的影响 将在体内使用表达不同种类的酿酒酵母菌株进行检测 CTP水平。CTP水平将通过表达 URA7基因，编码CTP合成酶。信使核糖核酸丰度和蛋白质 作为回应，将检测磷脂生物合成酶的水平 到不同级别的CTP。植物体内酶活性的生化调控 还将审查对CTP的反应。因为细胞CTP水平必须是 受CTP合成酶的表达和/或活性控制。这 应用程序还将解决CTP合成酶活性是否受 已知的调节磷脂合成的因素。CTP合成酶将是 通过分子遗传和/或生化方法纯化的 制备用于调节的抗CTP合成酶抗体的目的 学习。纯化的酶将根据其特性进行表征 基本的酶学和动力学性质。CTP的调控 通过cAMP依赖的蛋白激酶磷酸化的合成酶 在体内和体外进行检测。将从以下位置获取的信息 这些研究应该对其他方面的工作有很大的帮助和补充 磷脂代谢领域的研究人员以及 致力于核苷酸新陈代谢。

In the yeast Saccharomyces cerevisiae, CTP is a precursor of all membrane phospholipids synthesized by the phosphatidylethanolamine methylation (primary) and CDP-ethanolamine- and CDP-choline-based (auxiliary) pathways. This grant application addresses the hypothesis that cellular levels of CTP regulate the synthesis of phospholipids by the primary and auxiliary pathways by controlling the expression and/or activities of the enzymes in these pathways. The effects of CTP on phospholipid synthesis will be examined in vivo using S. cerevisiae strains expressing various levels of CTP. CTP levels will be varied through the expression of the URA7 gene, which encodes CTP synthetase. The mRNA abundance and protein levels of phospholipid biosynthetic enzymes will be examined in response to various levels of CTP. Biochemical regulation of enzyme activities in response to CTP will also be examined. Since cellular CTP levels must be governed by the expression and/or activity of CTP synthetase. This application will also address if CTP synthetase activity is regulated by factors known to regulated phospholipid synthesis. CTP synthetase will be purified by molecular genetic and/or biochemical approaches for the purpose of preparing anti-CTP synthetase antibodies for regulation studies. The purified enzyme will be characterized with respect to its basic enzymological and kinetic properties. The regulation of CTP synthetase by cAMP-dependent protein kinase phosphorylation will be examined in vivo and in vitro. The information that will be obtained from these studies should be of great use to, and complement the work of other investigators in the field of phospholipid metabolism as well as those working on nucleotide metabolism.