Cloning of a functional vesicular GABA and glycine transporter by screening of genome databases.
The unc-47 locus of Caenorhabditis elegans has been suggested to encode a synaptic vesicle GABA transporter. Here we used hydropathy plot analysis to identify a candidate vesicular GABA transporter in genomic sequences derived from a region of the physical map comprising unc-47. A mouse homologue was identified and cloned from EST database information. In situ hybridization in rat brain revealed codistribution with both GABAergic and glycinergic neuronal markers. Moreover, expression in COS-7 and PC12 cells induced an intracellular, glycine-sensitive GABA uptake activity. These observations, consistent with previous data on GABA and glycine uptake by synaptic vesicles, demonstrate that the mouse clone encodes a vesicular inhibitory amino acid transporter.
Sagné
C
C
CNRS ERS 575, Institut de Biologie Physico-Chimique, Paris, France.
El Mestikawy
S
S
Isambert
M F
MF
Hamon
M
M
Henry
J P
JP
Giros
B
B
Gasnier
B
B
D000595:Amino Acid Sequence; D026921:Amino Acid Transport Systems, Neutral; D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D003360:Cosmids; D016208:Databases, Factual; D050485:GABA Plasma Membrane Transport Proteins; D017238:Genes, Helminth; D050487:Glycine Plasma Membrane Transport Proteins; D006801:Humans; D017403:In Situ Hybridization; D008565:Membrane Proteins / Q000235:genetics*; D026901:Membrane Transport Proteins; D051379:Mice; D008969:Molecular Sequence Data; D027361:Organic Anion Transporters; D051381:Rats; D016415:Sequence Alignment; D014018:Tissue Distribution; D050495:Vesicular Inhibitory Amino Acid Transport Proteins
D026921:Amino Acid Transport Systems, Neutral; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D050485:GABA Plasma Membrane Transport Proteins; D050487:Glycine Plasma Membrane Transport Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D027361:Organic Anion Transporters; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; C493420:unc-47 protein, C elegans