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Elucidation of headgroup-specific functions of membrane phospholipids by molecular genetic approach.

通过分子遗传学方法阐明膜磷脂的头基特异性功能。

基本信息

批准号：
62470118
负责人：
SHIBUYA Isao
金额：
$ 4.16万
依托单位：
SAITAMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

Analyses by molecular genetic means have been carried out to elucidate the headgroup-specific functions of membrane phospholipids in Escherichia coli, Saccharomyces cerevisiae, and mouse FM3A cells.The E. coli cls gene was identified to encode cardiolipin synthase by purification and characterization of its protein product. Construction of null cls mutants indicated the non-essential nature of the cls gene and cardiolipin synthase, whereas cardiolipin, which is also formed by phosphatidylserine synthase, is most probably indispensable in E. coli. Discovery of the lethal effect of the pgsA3 mutation that encodes a defective phosphatidylglycerophosphate synthase, as well as of its suppression by a defect in the major outer membrane lipoptotein, indicated the essential nature and a measure of the necessary amount of phosphatidylglycerol for the survival of E. coli. A detailed analysis of pss-1 mutants which harbor a temperature-sensitive phosphatidylserine synthase showed the essential na … More ture on the matrix level and a measure of necessary amount of phosphatidylethanolamine.The S. cerevisiae CHO1 gene that encodes phosphatidylserine synthase was cloned, sequenced, and its primary product, which differs in size from the mature active form, was identified and characterized. A null CHO1 mutant was constructed to analyze the biological significance of phosphatidylserine. A phenotypic complementation of the phosphatidylserine deficiency with an increase in phosphatidylinositol and a novel route for phosphatidylethanolamine synthesis were observed. In S. cerevisiae, intracellular localization of the enzymes involved in phospholipid biosynthesis has been examined, and the enzymatic properties and regulatory features of mitochondrial phosphatidylglycerophosphate synthase and phosphatidylserine decarboxylase were determined.From mouse EM3A cells, mutants that require phosphatidylinositol for growth have been isolated and their phospholipid synthesis and myo-inositol metabolism were examined. The involvement of phosphatidylinositol in the regulation of intracellular myo-inositol level and phosphatidylglycerol synthesis have been indicated. Less

用分子遗传学方法分析了大肠杆菌、酿酒酵母和小鼠FM 3A细胞膜磷脂的头基特异性功能。通过对其蛋白产物的纯化和表征，鉴定大肠杆菌cls基因编码心磷脂合酶。cls缺失突变体的构建表明cls基因和心磷脂合酶的非必需性质，而心磷脂，也是由磷脂酰丝氨酸合酶形成的，在E.杆菌发现编码缺陷型磷脂酰甘油磷酸合酶的pgsA 3突变的致死效应，以及其被主要外膜脂蛋白缺陷抑制的效应，表明了E.杆菌对pss-1突变体进行的详细分析表明，温度敏感的磷脂酰丝氨酸合酶的必需na ...更多信息基质水平上的真实性和必需量的磷脂酰乙醇胺的测量。对编码磷脂酰丝氨酸合成酶的酿酒酵母CHO 1基因进行了克隆、测序，并鉴定和表征了其大小与成熟活性形式不同的初级产物。构建了CHO 1缺失突变体，以分析磷脂酰丝氨酸的生物学意义。磷脂酰丝氨酸缺乏症的表型互补与磷脂酰肌醇的增加和磷脂酰乙醇胺合成的新途径进行了观察。In S.从小鼠EM 3A细胞中分离出了需要磷脂酰肌醇生长的突变体，并对它们的磷脂合成和肌醇代谢进行了研究。磷脂酰肌醇参与细胞内肌醇水平和磷脂酰甘油合成的调节。少