Identification of 70 amino acids important for GABA(C) receptor rho1 subunit assembly

gamma-Aminobutyric acid (GABA) is the most important inhibitory neurotransmitter in the mammalian central nervous system and gates at least three subclasses of receptors, termed GABA(A), GABA(B) and GABA(C). Accumulating evidence indicates that GABA(C) receptors are composed exclusively of rho subunits. The N-terminal half of the rho subunits has been shown to mediate formation of homo- and heterooligomeric GABA(C) receptors. In this study, we searched for specific sequences within the N-terminus of the rho1 subunit involved in the assembly process. Assembly sequences were localized to a 128-amino acid region by deletion of progressively larger regions of a chimeric rho1beta1 subunit previously shown to disrupt rho1 and rho2 assembly. To confirm this observation, a series of GABA(A) receptor beta subunit chimeras containing different regions of the rho1 N-terminus were tested for interference with rho1 and rho2 subunit assembly into functional GABA receptors. Transfer of 70 residues within the 128 amino acid region to the beta1 subunit created a chimera that disrupted rho1, but not rho2, assembly into functional receptors. These observations refine the location of signals involved in rho1 subunit assembly, and suggest that different signals exist for the formation of rho1 homooligomeric and rho1/rho2 heterooligomeric GABA(C) receptors.