REGULATION OF CHOLINE KINASE ISOZYMES

磷脂酰胆碱是真核细胞的重要组成成分 膜，分泌脂蛋白和信号转导途径。其 生物合成部分由生物合成的第一种酶调节。 磷脂酰胆碱途径，胆碱激酶。的目的 该提案中概述的研究旨在了解 胆碱激酶活性的调节。有待检验的假设是， 该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产生的， 另一些则由不同的基因编码。这些研究将集中在两个 有机体为了扩展以前对哺乳动物胆碱激酶的研究， 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C。elegans将是 采用具体目标是：（1）开发分子工具， 鼠胆碱激酶这些工具将包括cDNA，抗体， 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激酶同工酶 表达ras癌基因。3)识别和表征小鼠 胆碱激酶基因4)表达和表征假定的胆碱 C.优雅5)研究胆碱的模式 C.优雅该研究将提供 关于磷脂酰胆碱合成的基础知识 调节，以及胆碱激酶如何参与其他细胞 功能协调发展的

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.