Cloning and expression of the MEP1 gene encoding an ammonium transporter in Saccharomyces cerevisiae.
In Saccharomyces cerevisiae, the transport of ammonium across the plasma membrane for use as a nitrogen source is mediated by at least two functionally distinct transport systems whose respective encoding genes are called MEP1 and MEP2. Mutations in the MEP2 gene affect high affinity, low capacity ammonium transport while mutations in the MEP1 gene disrupt a lower affinity, higher capacity system. In this work, the MEP1 gene has been cloned and sequenced and its expression analyzed. The predicted amino acid sequence reveals a highly hydrophobic, 54 kDa protein with 10 or 11 putative membrane-spanning regions. The predicted Mep1p protein shares high sequence similarity with several bacterial proteins of unknown function, notably the product of the nitrogen-regulated nrgA gene of Bacillus subtilis, and with that of a partial cDNA sequence derived from Caenorhabditis elegans. The Mep1p and related proteins appear to define a new family of transmembrane proteins evolutionarily conserved in at least bacteria, fungi and animals. The MEP1 gene is most highly expressed when the cells are grown on low concentrations of ammonium or on 'poor' nitrogen sources like urea or proline. It is down-regulated, on the other hand, when the concentration of ammonium is high or when other 'good' nitrogen sources like glutamine or asparagine are supplied in the culture medium. The overall properties of Mep1p indicate that it is a transporter of ammonium. Its main function appears to be to enable cells grown under nitrogen-limiting conditions to incorporate ammonium present at relatively low concentrations in the growth medium.
Marini
A M
AM
Laboratoire de Physiologie Cellulaire et de Génétique des Levures, Université Libre de Bruxelles, Belgium.
Vissers
S
S
Urrestarazu
A
A
André
B
B
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D027682:Cation Transport Proteins; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal / Q000235:genetics*; D017136:Ion Transport; D008744:Methylamines / Q000378:metabolism; D008969:Molecular Sequence Data; D000644:Quaternary Ammonium Compounds / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D000596:Amino Acids; D002352:Carrier Proteins; D027682:Cation Transport Proteins; D005656:Fungal Proteins; C088951:MEP1 protein, S cerevisiae; D008744:Methylamines; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; C027451:methylamine