REGULATION OF CHOLINE KINASE ISOZYMES

卵磷脂是真核细胞的重要组成部分 膜、分泌脂蛋白和信号转导通路。它的 生物合成部分由生物合成的第一种酶调节。 磷脂酰胆碱、胆碱激酶的途径。该计划的目标是 这项建议中概述的研究是为了了解 胆碱激酶活性的调节。需要检验的假设是 这种调节涉及多种同工酶的差异表达。 胆碱激酶，其中一些是通过选择性剪接和 其他基因编码在不同的基因中。这些研究将集中在两个方面 有机体。为了扩大以往对哺乳动物胆碱激酶的研究，小鼠 将使用组织和细胞系。开始使用分子遗传学 研究这些酶的方法，无脊椎动物线虫将是 使用。具体目标是i)开发分子工具来研究 小鼠胆碱酯酶。这些工具将包括cDNA、抗体和 重组蛋白。2)研究胆碱的表达模式 成年和胚胎小鼠组织和小鼠细胞系中的激酶同工酶 表达ras癌基因。3)鉴定和鉴定小鼠 胆碱激酶基因。4)表达和鉴定可能的胆碱 线虫的激酶同工酶。5)调查胆碱的类型 蛋白激酶在线虫中的表达。拟议的研究将提供 关于磷脂酰胆碱合成的基础知识 以及胆碱激酶可能如何参与其他细胞 功能。

Phosphatidylcholine is an important constituent of eukaryotic cell membranes, secreted lipoproteins, and signal transduction pathways. Its biosynthesis is regulated in part by the first enzyme of the biosynthetic pathway for phosphatidylcholine, choline kinase. The objective of the research outlined in this proposal is to understand the mechanism of regulation of choline kinase activity. The hypothesis to be tested is that the regulation involves differential expression of multiple isozymes of choline kinase, some of which are generated by alternative splicing and others are encoded in separate genes. These studies will focus on two organisms. To extend previous studies on mammalian choline kinase, mouse tissues and cell lines will be used. To begin to use a molecular genetic approach for studying these enzymes, the invertebrate C. elegans will be used. The specific aims are I) To develop molecular tools to investigate murine choline kinase. These tools will include cDNAs, antibodies, and recombinant protein. 2) To investigate patterns of expression of choline kinase isozymes in adult and embryonic mouse tissues and mouse cell lines expressing the ras oncogene. 3) To identify and characterize murine choline kinase genes. 4) To express and characterize putative choline kinase isozymes of C. elegans. 5) To investigate patterns of choline kinase expression in C. elegans. The proposed research will provide fundamental knowledge about how the synthesis of phosphatidylcholine is regulated, and about how choline kinase may be involved in other cellular functions.