Identification of a peroxisomal ATP carrier required for medium-chain fatty acid beta-oxidation and normal peroxisome proliferation in Saccharomyces cerevisiae

Mol Cell Biol. 2001 Jul;21(13):4321-9. doi: 10.1128/MCB.21.13.4321-4329.2001.

Abstract

We have characterized the role of YPR128cp, the orthologue of human PMP34, in fatty acid metabolism and peroxisomal proliferation in Saccharomyces cerevisiae. YPR128cp belongs to the mitochondrial carrier family (MCF) of solute transporters and is localized in the peroxisomal membrane. Disruption of the YPR128c gene results in impaired growth of the yeast with the medium-chain fatty acid (MCFA) laurate as a single carbon source, whereas normal growth was observed with the long-chain fatty acid (LCFA) oleate. MCFA but not LCFA beta-oxidation activity was markedly reduced in intact ypr128cDelta mutant cells compared to intact wild-type cells, but comparable activities were found in the corresponding lysates. These results imply that a transport step specific for MCFA beta-oxidation is impaired in ypr128cDelta cells. Since MCFA beta-oxidation in peroxisomes requires both ATP and CoASH for activation of the MCFAs into their corresponding coenzyme A esters, we studied whether YPR128cp is an ATP carrier. For this purpose we have used firefly luciferase targeted to peroxisomes to measure ATP consumption inside peroxisomes. We show that peroxisomal luciferase activity was strongly reduced in intact ypr128cDelta mutant cells compared to wild-type cells but comparable in lysates of both cell strains. We conclude that YPR128cp most likely mediates the transport of ATP across the peroxisomal membrane.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Carrier Proteins / chemistry
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Fractionation
  • Fatty Acids / metabolism*
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Genes, Reporter / genetics
  • Glucose / metabolism
  • Humans
  • Immunoblotting
  • Lauric Acids / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mitochondria / chemistry
  • Mitochondria / metabolism
  • Nucleotide Transport Proteins*
  • Oleic Acid / metabolism
  • Oxidation-Reduction
  • Peroxisomes / chemistry
  • Peroxisomes / metabolism*
  • Peroxisomes / ultrastructure
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / physiology*
  • Saccharomyces cerevisiae / ultrastructure
  • Saccharomyces cerevisiae Proteins*

Substances

  • ANT1 protein, S cerevisiae
  • Carrier Proteins
  • Fatty Acids
  • Fungal Proteins
  • Lauric Acids
  • Membrane Proteins
  • Nucleotide Transport Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • peroxisomal membrane protein 34
  • lauric acid
  • Oleic Acid
  • Adenosine Triphosphate
  • Luciferases
  • Glucose