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Isolation and characterization of cultured mammalian cell mutants defective in phospholipid biosynthesis

磷脂生物合成缺陷的培养哺乳动物细胞突变体的分离和表征

基本信息

批准号：
62571004
负责人：
NISHIJIMA Masahiro
金额：
$ 1.22万
依托单位：
National Institute of Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
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

(1) We cloned two CHO cell genes that complement the mutation of PSA-3, a phosphatidylserine auxotrophic mutant of CHO cell line. (2) We examined the effects of the modification of membrane phospholipids on the proliferation of the sindbis virus in PSA-3 cells and found that when PSA-3 cells are grown without phosphatidylserine, the binding and internalization of the virus occur normally but the yield of virions and viral RNA synthesis greatly decreased. These results indicate that cellular phosphatidylserine and/or phosphatidylethanolamine participate in a certain step of sindbis virus infection, after the internalization of virus particles, but before penetration of the viral nucleocapsids into the cytoplasm. (3) We obtained a CHO cell mutant (# 29) which is defective in the regulation of phosphatidylserine biosynthesis. In this mutant cells, phosphatidylserine bisynthesis was elevated about two-fold and was remarkably resistant to the inhibition by exogenous phosphatidylserine as compared to the parental cells. (4) We found that CHO cells have two kinds of inositol transport systems, namely, sodium-dependent and sodium-independent systems. We obtained CHO cell mutants defective in sodium-dependent inositol transport system. (5) We isolated CHO cell mutants defective in glucose transport. In one of the mutants (GTS-31), the level of glucose transporter was reduced by 80% as compared to the parental cells.

(1)我们克隆了两个CHO细胞基因，补充PSA-3，CHO细胞系的磷脂酰丝氨酸营养缺陷型突变体的突变。(2)我们研究了膜磷脂的修饰对PSA-3细胞中辛德毕斯病毒增殖的影响，发现当PSA-3细胞在没有磷脂酰丝氨酸的情况下生长时，病毒的结合和内化正常发生，但病毒粒子的产量和病毒RNA的合成大大降低。这些结果表明，细胞磷脂酰丝氨酸和/或磷脂酰乙醇胺参与辛德毕斯病毒感染的某个步骤，在病毒颗粒的内化之后，但在病毒核衣壳渗透到细胞质之前。(3)我们获得了CHO细胞突变体（#29），其在磷脂酰丝氨酸生物合成的调节中有缺陷。在这种突变体细胞中，磷脂酰丝氨酸双合成升高约两倍，并显着抵抗外源磷脂酰丝氨酸的抑制相比，亲本细胞。(4)我们发现CHO细胞具有两种肌醇转运系统，即钠依赖型和非钠依赖型。我们获得了钠依赖性肌醇转运系统缺陷的CHO细胞突变体。(5)我们分离了葡萄糖转运缺陷的CHO细胞突变体。在一个突变体（GTS-31）中，与亲本细胞相比，葡萄糖转运蛋白的水平降低了80%。