Dip5p mediates high-affinity and high-capacity transport of L-glutamate and L-aspartate in Saccharomyces cerevisiae.
Genes encoding homologues of known amino-acid permeases were deleted in a strain also deficient in the general amino-acid permease. The uptake capacity of the mutants was investigated for several L-alpha-amino acids. Deletion of a gene denoted DIP5 results in the loss of L-aspartate and L-glutamate uptake. The dip5 mutation caused a several hundred-fold reduction of uptake of the two amino acids, both in cells grown on proline as a nitrogen source and in cells grown on ammonium. DIP5-dependent uptake of L-aspartate and L-glutamate was somewhat lower in ammonium-grown cells than in proline-grown cells. Transcriptional regulation is at least partially responsible for this difference, as shown by assaying the DIP5 promoter fused to lacZ. This suggests that the promoter is subject to nitrogen catabolite repression. Transport of a few other amino acids was moderately affected by dip5 but was not competed by L-aspartate in the DIP5 parental strain; transport of these amino acids is therefore unlikely to be mediated by Dip5p. Our results suggest that DIP5 encodes an amino-acid permease with a high transport capacity and a high affinity for L-glutamate and L-aspartate, with a Kt of about 50 microM for both.
Regenberg
B
B
Department of Yeast Genetics, Carlsberg Laboratory, DK-2500 Copenhagen Valby, Denmark. breg@crc.dk
Holmberg
S
S
Olsen
L D
LD
Kielland-Brandt
M C
MC
D026905:Amino Acid Transport Systems; D026922:Amino Acid Transport Systems, Acidic; D000596:Amino Acids / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism*; D001692:Biological Transport; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D005800:Genes, Fungal / Q000235:genetics; D007700:Kinetics; D026901:Membrane Transport Proteins / Q000378:metabolism*; D009584:Nitrogen / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D017384:Sequence Deletion
D026905:Amino Acid Transport Systems; D026922:Amino Acid Transport Systems, Acidic; D000596:Amino Acids; C503357:DIP5 protein, S cerevisiae; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D001224:Aspartic Acid; D009584:Nitrogen