Phosphatidyl-N-methylethanolamine methyltransferase mutants of Saccharomyces cerevisiae were isolated. Genetic analysis showed that phosphatidylethanolamine methyltransferase and phosphatidyl-N-methylethanolamine methyltransferase are coded for by separate genes. Phosphatidyl-N-methylethanolamine methyltransferase activity and phosphatidyl-N,N-dimethylethanolamine methyltransferase activity appeared to be catalyzed by the same enzyme. 2. Phosphatidyl-N-methylethanolamine methyltransferase was found to be repressed by myo-inositol and choline. Both myo-inositol and choline at concentrations of 10 micrograms/ml were required for repression. The decreased enzyme level was restored by the removal of myo-inositol or choline or both. 3. Both myo-inositol and choline were required for the maximum repression of phosphatidylethanolamine methyltransferase in wild-type cells. In contrast, choline was not required for the repression of the enzyme in mutant strain 172. This was due to a single nuclear gene mutation in the genome of strain 172. 4. The activity of the phosphatidylethanolamine methylation pathway in cells decreased with time on incubation of cells with myo-inositol and choline, myo-Inositol could not be replaced by other structurally related compounds, such as scyllo-inositol or mannitol. 5. The physiological significance of the repression of the phosphatidylethanolamine methylation pathway is discussed with respect to the mechanism for maintaining the contents of phosphatidylethanolamine and phosphatidylcholine at normal levels.
摘要
分离出了酿酒酵母的磷脂酰 - N - 甲基乙醇胺甲基转移酶突变体。遗传分析表明,磷脂酰乙醇胺甲基转移酶和磷脂酰 - N - 甲基乙醇胺甲基转移酶由不同基因编码。磷脂酰 - N - 甲基乙醇胺甲基转移酶活性和磷脂酰 - N,N - 二甲基乙醇胺甲基转移酶活性似乎由同一种酶催化。2. 发现磷脂酰 - N - 甲基乙醇胺甲基转移酶受肌醇和胆碱的抑制。肌醇和胆碱浓度均为10微克/毫升时才会产生抑制作用。去除肌醇或胆碱或两者后,酶水平降低的情况得以恢复。3. 野生型细胞中,肌醇和胆碱都是磷脂酰乙醇胺甲基转移酶最大程度抑制所必需的。相比之下,突变株172中该酶的抑制不需要胆碱。这是由于172菌株基因组中的一个单细胞核基因突变所致。4. 细胞与肌醇和胆碱一起孵育时,细胞中磷脂酰乙醇胺甲基化途径的活性随时间下降。肌醇不能被其他结构相关化合物如 scyllo - 肌醇或甘露醇替代。5. 就维持磷脂酰乙醇胺和磷脂酰胆碱含量在正常水平的机制而言,讨论了磷脂酰乙醇胺甲基化途径抑制的生理意义。