Mutagenesis of the putative sterol-sensing domain of yeast Niemann Pick C-related protein reveals a primordial role in subcellular sphingolipid distribution

J Cell Biol. 2004 Feb 16;164(4):547-56. doi: 10.1083/jcb.200310046.

Abstract

Lipid movement between organelles is a critical component of eukaryotic membrane homeostasis. Niemann Pick type C (NP-C) disease is a fatal neurodegenerative disorder typified by lysosomal accumulation of cholesterol and sphingolipids. Expression of yeast NP-C-related gene 1 (NCR1), the orthologue of the human NP-C gene 1 (NPC1) defective in the disease, in Chinese hamster ovary NPC1 mutant cells suppressed lipid accumulation. Deletion of NCR1, encoding a transmembrane glycoprotein predominantly residing in the vacuole of normal yeast, gave no phenotype. However, a dominant mutation in the putative sterol-sensing domain of Ncr1p conferred temperature and polyene antibiotic sensitivity without changes in sterol metabolism. Instead, the mutant cells were resistant to inhibitors of sphingolipid biosynthesis and super sensitive to sphingosine and C2-ceramide. Moreover, plasma membrane sphingolipids accumulated and redistributed to the vacuole and other subcellular membranes of the mutant cells. We propose that the primordial function of these proteins is to recycle sphingolipids and that defects in this process in higher eukaryotes secondarily result in cholesterol accumulation.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biological Transport / physiology
  • CHO Cells
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism*
  • Cell Membrane / metabolism
  • Cricetinae
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / metabolism*
  • Molecular Sequence Data
  • Niemann-Pick C1 Protein
  • Niemann-Pick Diseases / genetics
  • Niemann-Pick Diseases / metabolism
  • Protein Structure, Tertiary
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Sequence Alignment
  • Sphingolipids / metabolism*
  • Sterols / metabolism*
  • Subcellular Fractions / chemistry
  • Subcellular Fractions / metabolism
  • Vesicular Transport Proteins

Substances

  • Carrier Proteins
  • Intracellular Signaling Peptides and Proteins
  • Membrane Glycoproteins
  • NCR1 protein, S cerevisiae
  • NPC1 protein, human
  • Niemann-Pick C1 Protein
  • Saccharomyces cerevisiae Proteins
  • Sphingolipids
  • Sterols
  • Vesicular Transport Proteins