title: The RXR-type endoplasmic reticulum-retention/retrieval signal of GABAB1 requires distant spacing from the membrane to function.
abstract: Functional gamma-aminobutyric acid type B (GABA(B)) receptors are normally only observed upon coexpression of GABA(B1) with GABA(B2) subunits. A C-terminal arginine-based endoplasmic reticulum (ER) retention/retrieval signal, RSRR, prevents escape of unassembled GABA(B1) subunits from the ER and restricts surface expression to correctly assembled heteromeric receptors. The RSRR signal in GABA(B1) is proposed to be shielded by C-terminal coiled-coil interaction of the GABA(B1) with the GABA(B2) subunit. Here, we investigated whether the RSRR motif in GABA(B1) remains functional when grafted to ectopic sites. We found that the RSRR signal in GABA(B1) is inactive in any of the three intracellular loops but remains functional when moved within the distal zone of the C-terminal tail. C-terminal deletions that position the RSRR signal closer to the plasma membrane drastically reduce its effectiveness, supporting that proximity to the membrane restricts access to the RSRR motif. Functional ectopic RSRR signals in GABA(B1) are efficiently inactivated by the GABA(B2) subunit in the absence of coiled-coil dimerization, supporting that coiled-coil interaction is not critical for release of the receptor complex from the ER. The data are consistent with a model in which removal of RSRR from its active zone rather than its direct shielding by coiled-coil dimerization triggers forward trafficking. Because arginine-based intracellular retention signals of the type RXR, where X represents any amino acid, are used to regulate assembly and surface transport of several multimeric complexes, such a mechanism may apply to other proteins as well.
authors.affiliation: Pharmazentrum, University of Basel, Department of Clinical-Biological Sciences, Institute of Physiology, Basel, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Gassmann
authors.forename: Corinne
authors.initials: C
authors.lastname: Haller
authors.forename: Yanick
authors.initials: Y
authors.lastname: Stoll
authors.forename: Said
authors.initials: S
authors.lastname: Abdel Aziz
authors.forename: Barbara
authors.initials: B
authors.lastname: Biermann
authors.forename: Johannes
authors.initials: J
authors.lastname: Mosbacher
authors.forename: Klemens
authors.initials: K
authors.lastname: Kaupmann
authors.forename: Bernhard
authors.initials: B
authors.lastname: Bettler
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000235:genetics / Q000502:physiology*; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000502:physiology*; D006801:Humans; D018080:Receptors, GABA-B / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D047488:Retinoid X Receptors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D018080:Receptors, GABA-B; D047488:Retinoid X Receptors