Molecular characterization of carnitine-dependent transport of acetyl-CoA from peroxisomes to mitochondria in Saccharomyces cerevisiae and identification of a plasma membrane carnitine transporter, Agp2p.
In Saccharomyces cerevisiae, beta-oxidation of fatty acids is confined to peroxisomes. The acetyl-CoA produced has to be transported from the peroxisomes via the cytoplasm to the mitochondrial matrix in order to be degraded to CO(2) and H(2)O. Two pathways for the transport of acetyl-CoA to the mitochondria have been proposed. The first involves peroxisomal conversion of acetyl-CoA into glyoxylate cycle intermediates followed by transport of these intermediates to the mitochondria. The second pathway involves peroxisomal conversion of acetyl-CoA into acetylcarnitine, which is subsequently transported to the mitochondria. Using a selective screen, we have isolated several mutants that are specifically affected in the second pathway, the carnitine-dependent acetyl-CoA transport from the peroxisomes to the mitochondria, and assigned these CDAT mutants to three different complementation groups. The corresponding genes were identified using functional complementation of the mutants with a genomic DNA library. In addition to the previously reported carnitine acetyl-CoA transferase (CAT2), we identified the genes for the yeast orthologue of the human mitochondrial carnitine acylcarnitine translocase (YOR100C or CAC) and for a transport protein (AGP2) required for carnitine transport across the plasma membrane.
van Roermund
C W
CW
University of Amsterdam, Academic Medical Centre, Department of Clinical Chemistry, Emma Children's Hospital, The Netherlands.
Hettema
E H
EH
van den Berg
M
M
Tabak
H F
HF
Wanders
R J
RJ
D000105:Acetyl Coenzyme A / Q000378:metabolism*; D026905:Amino Acid Transport Systems; D001692:Biological Transport / Q000235:genetics; D002331:Carnitine / Q000378:metabolism*; D002333:Carnitine Acyltransferases / Q000378:metabolism; D002332:Carnitine O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry*; D003001:Cloning, Molecular; D005656:Fungal Proteins; D005800:Genes, Fungal; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D016253:Microscopy, Immunoelectron; D008928:Mitochondria / Q000378:metabolism*; D009154:Mutation; D019301:Oleic Acid / Q000378:metabolism; D020675:Peroxisomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D027981:Symporters
C402286:Agp2 protein, S cerevisiae; D026905:Amino Acid Transport Systems; D002352:Carrier Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; D027981:Symporters; D019301:Oleic Acid; D000105:Acetyl Coenzyme A; D002333:Carnitine Acyltransferases; D002332:Carnitine O-Acetyltransferase; D002331:Carnitine