C-terminal interaction is essential for surface trafficking but not for heteromeric assembly of GABA(b) receptors.
Assembly of fully functional GABA(B) receptors requires heteromerization of the GABA(B(1)) and GABA(B(2)) subunits. It is thought that GABA(B(1)) and GABA(B(2)) undergo coiled-coil dimerization in their cytoplasmic C termini and that assembly is necessary to overcome GABA(B(1)) retention in the endoplasmatic reticulum (ER). We investigated the mechanism underlying GABA(B(1)) trafficking to the cell surface. We identified a signal, RSRR, proximal to the coiled-coil domain of GABA(B(1)) that when deleted or mutagenized allows for surface delivery in the absence of GABA(B(2)). A similar motif, RXR, was recently shown to function as an ER retention/retrieval (ERR/R) signal in K(ATP) channels, demonstrating that G-protein-coupled receptors (GPCRs) and ion channels use common mechanisms to control surface trafficking. A C-terminal fragment of GABA(B(2)) is able to mask the RSRR signal and to direct the GABA(B(1)) monomer to the cell surface, where it is functionally inert. This indicates that in the heteromer, GABA(B(2)) participates in coupling to the G-protein. Mutagenesis of the C-terminal coiled-coil domains in GABA(B(1)) and GABA(B(2)) supports the possibility that their interaction is involved in shielding the ERR/R signal. However, assembly of heteromeric GABA(B) receptors is possible in the absence of the C-terminal domains, indicating that coiled-coil interaction is not necessary for function. Rather than guaranteeing heterodimerization, as previously assumed, the coiled-coil structure appears to be important for export of the receptor complex from the secretory apparatus.
Pagano
A
A
Novartis Pharma AG, Therapeutic Area Nervous System, CH-4002 Basle, Switzerland.
Rovelli
G
G
Mosbacher
J
J
Lohmann
T
T
Duthey
B
B
Stauffer
D
D
Ristig
D
D
Schuler
V
V
Meigel
I
I
Lampert
C
C
Stein
T
T
Prezeau
L
L
Blahos
J
J
Pin
J
J
Froestl
W
W
Kuhn
R
R
Heid
J
J
Kaupmann
K
K
Bettler
B
B
D020816:Amino Acid Motifs / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D019281:Dimerization; D004721:Endoplasmic Reticulum / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D019920:Photoaffinity Labels / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021122:Protein Subunits; D021381:Protein Transport / Q000502:physiology*; D018080:Receptors, GABA-B / Q000235:genetics / Q000378:metabolism*; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology
D019920:Photoaffinity Labels; D021122:Protein Subunits; D018080:Receptors, GABA-B; D018094:Receptors, Metabotropic Glutamate; D019204:GTP-Binding Proteins; D002118:Calcium