title: Mechanisms of endoplasmic-reticulum export of glycine transporter-1 (GLYT1).
abstract: The GLYT1 (glycine transporter-1) regulates both glycinergic and glutamatergic neurotransmission by controlling the reuptake of glycine at synapses. Trafficking to the cell surface of GLYT1 is critical for its function. In the present paper, by using mutational analysis of the GLYT1 C-terminal domain, we identified the evolutionarily conserved motif R(575)L(576)(X(8))D(585) as being necessary for ER (endoplasmic reticulum) export. This is probably due to its capacity to bind Sec24D, a component of the COPII (coatomer coat protein II) complex. This ER export motif was active when introduced into the related GLYT2 transporter but not in the unrelated VSVG (vesicular-stomatitis virus glycoprotein)-GLYT1 protein in which this motif was mutated but was not transported to the plasma membrane, although this effect was rescued by co-expressing these mutants with wild-type GLYT1. This behaviour suggests that GLYT1 might form oligomers along the trafficking pathway. Cross-linking assays performed in rat brain synaptosomes and FRET (fluorescence resonance energy transfer) microscopy in living cells confirmed the existence of GLYT1 oligomers. In summary, we have identified a motif involved in the ER exit of GLYT1 and, in analysing the influence of this motif, we have found evidence that oligomerization is important for the trafficking of GLYT1 to the cell surface. Because this motif is conserved in the NSS (sodium- and chloride-dependent neurotransmitter transporter) family, it is possible that this finding could be extrapolated to other related transporters.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Facultad de Ciencias, Universidad Autónoma de Madrid, 28049 Madrid, Spain.
authors.forename: Enrique
authors.initials: E
authors.lastname: Fernández-Sánchez
authors.forename: F Javier
authors.initials: FJ
authors.lastname: Díez-Guerra
authors.forename: Beatriz
authors.initials: B
authors.lastname: Cubelos
authors.forename: Cecilio
authors.initials: C
authors.lastname: Giménez
authors.forename: Francisco
authors.initials: F
authors.lastname: Zafra
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005786:Gene Expression Regulation; D050487:Glycine Plasma Membrane Transport Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D021381:Protein Transport; D051381:Rats; D016415:Sequence Alignment; D033921:Vesicular Transport Proteins / Q000378:metabolism
chemical_list: D050487:Glycine Plasma Membrane Transport Proteins; D033921:Vesicular Transport Proteins