Chinese hamster ovary cell mutant with defective down-regulation of low density lipoprotein receptors.
A monensin-resistant clone (Monr-31) shows a related series of differences from its parental Chinese hamster ovary (CHO) cell line in the cellular response to several ligands. The uptake and metabolism of low density lipoprotein (LDL) in the mutant cells are defective. Accumulation of fluorescent-labeled LDL as well as internalization and degradation of 125I-LDL are greatly reduced in Monr-31 cells. The receptor number for LDL on the cell surface of Monr-31 is about one-third that for CHO cells, but affinity constants for both cell lines are similar. Electrophoretic analysis shows a slightly reduced molecular weight of LDL receptor in Monr-31 cells in comparison to that in CHO cells. The internalization index (internalization plus degradation per binding) of LDL of the mutant is about one-half that of CHO cells, suggesting a failure of internalization of LDL as well as LDL binding. Hybrids (hyb-1, -2, and -3) between CHO and Monr-31 cells show LDL binding and LDL internalization activities comparable to that of CHO cells, suggesting that the altered LDL response in Monr-31 cells is recessive. Addition of exogenous LDL to culture medium down-regulates the LDL receptor activity of CHO, hyb-2, and hyb-3 cells, whereas no such down-regulation is seen in Monr-31 cells. Probably as a result of the failure of down-regulation, the prominent inhibition of sterol synthesis from acetate and 3-hydroxy-3-methylglutaryl-coenzyme A reductase observed in CHO cells is scarcely detectable in Monr-31 cells. As a correlated result, sterol synthesis from acetate is 6-fold higher in the mutant. The failure of down-regulation of LDL receptors in Monr-31 cells is discussed in relation to the altered binding and internalization of LDL.
D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D006224:Cricetinae; D004351:Drug Resistance; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006822:Hybrid Cells / Q000378:metabolism; D006888:Hydroxycholesterols / Q000494:pharmacology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008077:Lipoproteins, LDL / Q000378:metabolism* / Q000494:pharmacology; D008970:Molecular Weight; D008985:Monensin / Q000494:pharmacology; D009154:Mutation; D010053:Ovary; D011973:Receptors, LDL / Q000235:genetics* / Q000378:metabolism; D013261:Sterols / Q000096:biosynthesis
D006888:Hydroxycholesterols; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D013261:Sterols; C007997:25-hydroxycholesterol; D008985:Monensin; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases