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Mammalian cell mutant requiring cholesterol and unsaturated fatty acid for growth

, : Mammalian cell mutant requiring cholesterol and unsaturated fatty acid for growth. Proceedings of the National Academy of Sciences of the United States of America 75(11): 5452-5456

A mutant requiring both cholesterol and oleate for growth was isolated from mutagenized Chinese hamster ovary cells. By comparison with wild-type cells, sterol and unsaturated fatty acid biosynthetic activities in the mutant cells grown in fetal calf serum medium appear to be nearly intact. Whole-cell radioactive acetate, mevalonate, dihydrolanosterol, and stearate incorporation studies show that sterol synthesis from acetate, lanosterol demethylation, and fatty acid desaturation are defective in the mutant cells grown in delipidated serum medium. In vitro enzyme assays with crude cell extracts demonstrated that .beta.-hydroxy-.beta.-methylglutaryl-CoA reductase is not induced in the mutant. These experiments were substantiated by GLC analyses which showed the sterol content and the percentage unsaturated fatty acids in mutant cells to be drastically reduced when the cells are grown in delipidated serum medium. A spontaneous revertant exhibiting prototrophic growth in lipid-free medium was isolated from 50 .times. 106 mutant cells. All 3 defects in this revertant reverted back in parallel, which suggests that these 3 biosynthetic activities may be controlled by a common regulatory mechanism.


PMID: 281693

DOI: 10.2307/68649

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