Complementation of mutation in Acyl-CoA:cholesterol acyltransferase (ACAT) fails to restore sterol regulation in ACAT-defective sterol-resistant hamster cells

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

A previously described mutant line of Chinese hamster ovary cells, designated SRD-4, fails to synthesize cholesteryl esters, owing to a deficiency in the activity of acyl-CoA:cholesterol acyltransferase (ACAT). These cells also fail to suppress low density lipoprotein receptors or cholesterol synthesizing enzymes in the presence of 25-hydroxycholesterol. In the current studies we show that SRD-4 cells have three defects: 1) a point mutation in one allele at the ACAT locus that changes codon 265 from Ser to Leu, resulting in an inactive enzyme; 2) a silent allele at the other ACAT locus that does not produce detectable mRNA; and 3) a mutation, as yet undefined, that abolishes the ability of 25-hydroxycholesterol to inhibit the cleavage of both sterol regulatory element binding proteins (SREBP-1 and SREBP-2). Correction of the ACAT deficiency by transfection of a wild-type cDNA failed to restore inhibition of SREBP cleavage by 25- hydroxycholesterol, indicating that the ACAT deficiency and the sterol regulatory defect are caused by independent mutations. These data provide further insight into the interplay between ACAT activation and inhibition of SREBP cleavage by 25-hydroxycholesterol, and they indicate that these two processes can be disrupted independently by mutation.

Original languageEnglish (US)
Pages (from-to)14642-14648
Number of pages7
JournalJournal of Biological Chemistry
Volume271
Issue number24
DOIs
StatePublished - Jun 25 1996

    Fingerprint

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this