Sterol resistance in CHO cells traced to point mutation in SREBP cleavage-activating protein

Cell. 1996 Nov 1;87(3):415-26. doi: 10.1016/s0092-8674(00)81362-8.

Abstract

Through expression cloning we have isolated a cDNA-encoding SREBP cleavage-activating protein (SCAP), which regulates cholesterol metabolism by stimulating cleavage of transcription factors SREBP-1 and -2, thereby releasing them from membranes. The cDNA was isolated from Chinese hamster ovary cells with a dominant mutation that renders them resistant to sterol-mediated suppression of cholesterol synthesis and uptake. Sterol resistance was traced to a G-->A transition at codon 443 of SCAP, changing aspartic acid to asparagine. The D443N mutation enhances the cleavage-stimulating ability of SCAP and renders it resistant to inhibition by sterols. SCAP has multiple membrane-spanning regions, five of which resemble the sterol-sensing domain of HMG CoA reductase, an endoplasmic reticulum enzyme whose degradation is accelerated by sterols. SCAP appears to be a central regulator of cholesterol metabolism in animal cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CCAAT-Enhancer-Binding Proteins*
  • CHO Cells / drug effects
  • CHO Cells / metabolism*
  • Caenorhabditis elegans / genetics
  • Cholesterol / metabolism*
  • Cloning, Molecular
  • Cricetinae
  • Cricetulus
  • DNA, Complementary / genetics
  • DNA-Binding Proteins / metabolism*
  • Feedback
  • Gene Expression Regulation
  • Genes*
  • Genes, Reporter
  • Humans
  • Hydroxymethylglutaryl CoA Reductases / chemistry
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology*
  • Molecular Sequence Data
  • Nuclear Proteins / metabolism*
  • Receptors, LDL / biosynthesis*
  • Receptors, LDL / genetics
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Sterol Regulatory Element Binding Protein 1
  • Sterol Regulatory Element Binding Protein 2
  • Sterols / pharmacology*
  • Transcription Factors / metabolism
  • Transfection

Substances

  • CCAAT-Enhancer-Binding Proteins
  • DNA, Complementary
  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Nuclear Proteins
  • Receptors, LDL
  • SREBF1 protein, human
  • SREBF2 protein, human
  • SREBP cleavage-activating protein
  • Sterol Regulatory Element Binding Protein 1
  • Sterol Regulatory Element Binding Protein 2
  • Sterols
  • Transcription Factors
  • Cholesterol
  • Hydroxymethylglutaryl CoA Reductases

Associated data

  • GENBANK/D83782
  • GENBANK/U67060