Saccharomyces cerevisiae Aqr1 is an internal-membrane transporter involved in excretion of amino acids.
Excretion of amino acids by yeast cells was reported long ago but has not been characterized in molecular terms. It is typically favored by overproduction of the amino acid and/or impairment of its uptake. Here we describe the construction of a yeast strain excreting threonine and homoserine. Using this excretor strain, we then applied a reverse-genetics approach and found that the transporter encoded by the YNL065w/AQR1 gene, a protein thought to mediate H(+) antiport, is involved in homoserine and threonine excretion. Furthermore, overexpression of AQR1 led to increased excretion of several amino acids (alanine, aspartate, and glutamate) known to be relatively abundant in the cytosol. Transcription of the AQR1 gene is induced severalfold by a number of amino acids and appears to be under the negative control of Gcn4. An Aqr1-green fluorescent protein fusion protein is located in multiple internal membrane structures and appears to cycle continuously between these compartments and the plasma membrane. The Aqr1 sequence is significantly similar to the vesicular amine transporters of secretory vesicles of neuronal cells. We propose that Aqr1 catalyzes transport of excess amino acids into vesicles, which then release them in the extracellular space by exocytosis.
Velasco
Isabel
I
Physiologie Moléculaire de la Cellule, Université Libre de Bruxelles, Institut de Biologie et de Médecine Moléculaires, 12 rue des Pr. Jeener et Brachet, 6041 Gosselies, Belgium.
Tenreiro
Sandra
S
Calderon
Isabel L
IL
André
Bruno
B
D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis; D005089:Exocytosis; D015966:Gene Expression Regulation, Fungal; D005838:Genotype; D018698:Glutamic Acid / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006714:Homoserine / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism / Q000502:physiology*; D008744:Methylamines / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D027501:Monocarboxylic Acid Transporters; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D013912:Threonine / Q000378:metabolism; D013997:Time Factors; D001616:beta-Galactosidase / Q000378:metabolism
C454200:AQR1 protein, S cerevisiae; D000596:Amino Acids; D026901:Membrane Transport Proteins; D008744:Methylamines; D027501:Monocarboxylic Acid Transporters; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D013912:Threonine; D001224:Aspartic Acid; D018698:Glutamic Acid; D006714:Homoserine; C027451:methylamine; D001616:beta-Galactosidase; D000409:Alanine