Sequence of a functional invertebrate GABAA receptor subunit which can form a chimeric receptor with a vertebrate alpha subunit.
The sequence of an invertebrate GABAA receptor subunit is described. This was deduced from a cDNA which was isolated from the mollusc Lymnaea stagnalis and which corresponds to a transcript of extremely low abundance. The cDNA was isolated using short exonic sequences from part of the corresponding gene in combination with a variant of the polymerase chain reaction (PCR) known as RACE (rapid amplification of cDNA ends). The mature polypeptide has a predicted molecular weight of 54,569 Daltons and exhibits approximately 50% identity to vertebrate GABAA receptor beta subunits. The six intron-exon boundaries determined to date in the molluscan gene occur at the same relative positions as those found in vertebrate GABAA receptor genes. Functional expression, in Xenopus oocytes, of the molluscan cDNA alone results in the formation of GABA-activated chloride ion channels that have a finite open probability even in the absence of agonist. These GABA-evoked currents can be reversibly blocked by the vertebrate GABAA receptor antagonist bicuculline. Surprisingly, the molluscan beta subunit is capable of replacing vertebrate beta subunits in co-expression experiments with the bovine GABAA receptor alpha 1 subunit. These findings suggest that invertebrate GABAA receptors exist in vivo as hetero-oligomeric complexes.
Harvey
R J
RJ
MRC Molecular Neurobiology Unit, MRC Centre, Cambridge, UK.
Vreugdenhil
E
E
Zaman
S H
SH
Bhandal
N S
NS
Usherwood
P N
PN
Barnard
E A
EA
Darlison
M G
MG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002678:Chimera; D004247:DNA / Q000235:genetics; D008195:Lymnaea; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011963:Receptors, GABA-A / Q000235:genetics*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014981:Xenopus
D011963:Receptors, GABA-A; D004247:DNA