Evidence for coordinate expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase ad low density lipoprotein binding activity.
A Chinese hamster ovary cell mutant that requires both cholesterol and unsaturated fatty acids for growth (Limanek, J. S., Chin, J., and Chang, T. Y. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 5452-5456) has been further characterized with respect to its dependence on cholesterol. Upon removal of serum lipids from the growth medium, the activity of the important cholesterogenic enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and the low density lipoprotein (LDL) binding activity both increase significantly in the normal cell. Both these increases were much less in the mutant cell. Studies in vitro with NaF indicate that the differences in reductase activities between normal and mutant cells are not due to differences in activation by a dephosphorylation mechanism. Heat inactivation profiles and Km for HMG-CoA of both cell reductases were found to be identical, thus reducing the possibility that the mutant cell contains a mutation in the polypeptide chain of reductase. The fact that in lipid-deficient medium both reductase and LDL binding activities are low in the mutant strongly suggests that the expression of these activities is controlled in a coordinate manner. This conclusion is supported by parallel studies on a spontaneous revertant of the mutant in which the expression of reductase and LDL binding activities have both reverted to normal. These results indicate that the phenotypic abnormalities seen in the mutant are probably caused by a single mutation. A common factor is postulated to mediate this coordinate expression, and the function of such a factor is altered in the mutant cell.
D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D002784:Cholesterol / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus / Q000235:genetics; D003470:Culture Media; D005231:Fatty Acids, Unsaturated / Q000502:physiology; D005260:Female; D006888:Hydroxycholesterols / Q000494:pharmacology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism*; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008077:Lipoproteins, LDL / Q000378:metabolism*; D009154:Mutation; D010053:Ovary / Q000201:enzymology*
D003470:Culture Media; D005231:Fatty Acids, Unsaturated; D006888:Hydroxycholesterols; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008077:Lipoproteins, LDL; C007997:25-hydroxycholesterol; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases