Expression and regulation of human low-density lipoprotein receptors in Chinese hamster ovary cells

Abstract

Low-density lipoprotein (LDL), the major cholesterol transport component of human plasma, delivers cholesterol to mammalian cells via the LDL pathway of receptor-mediated endocytosis. LDL receptor activity and cholesterol biosynthesis are coordinately regulated by cholesterol-mediated feedback suppression. We have developed methods for the isolation of mutant and revertant (M.K., unpublished data) Chinese hamster ovary (CHO) cells having alterations in cholesterol biosynthesis and in the receptor-mediated endocytosis of LDL. The defective locus of one LDL receptor-negative CHO mutant (clone 7a-1) is apparently the structural gene for the LDL receptor (D. Kingsley, M. Segal and M.K., unpublished data). Here we have transfected 7a-1 cells with human DNA and selected colonies which synthesize functional human LDL receptors whose expression is regulated normally. This selection may prove useful for cloning genes required for the receptor-mediated endocytosis of LDL and other ligands and thus for elucidating the molecular defects responsible for familial hypercholesterolaemia, one of the most common genetic diseases in humans.