RNA editing of the GABA(A) receptor alpha3 subunit alters the functional properties of recombinant receptors.
RNA editing provides a post-transcriptional mechanism to increase structural heterogeneity of gene products. Recently, the alpha3 subunit of the GABAA receptors has been shown to undergo RNA editing. As a result, a highly conserved isoleucine residue in the third transmembrane domain is replaced with a methionine. To determine the effect of this structural change on receptor function, we compared the GABA sensitivity, pharmacological properties and macroscopic kinetics of recombinant receptors containing either the edited or unedited forms of the alpha3 subunit along with beta3 and gamma2L. Editing substantially altered the GABA sensitivity and deactivation rate of the receptors, with the unedited form showing a lower GABA EC50 and slower decay. Comparable effects were observed with a mutation at the homologous location in the alpha1 subunit, suggesting a common role for this site in regulation of channel gating. Except for the response to GABA, the pharmacological properties of the receptor were unaffected by editing, with similar enhancement by a variety of modulators. Since RNA editing of the alpha3 subunit increases through development, our findings suggest that GABAergic neurotransmission may be more effective early in development, with greater GABA sensitivity and slower decay rates conferred by the unedited alpha3 subunit.
Nimmich
Mitchell L
ML
Department of Pharmacology, Physiology and Neuroscience, University of South Carolina School of Medicine, Columbia 29208, USA.
Heidelberg
Laura S
LS
Fisher
Janet L
JL
D000595:Amino Acid Sequence; D002461:Cell Line, Transformed; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D018682:GABA Agents / Q000494:pharmacology; D006801:Humans; D007532:Isoleucine / Q000235:genetics; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008715:Methionine / Q000235:genetics; D009154:Mutation / Q000235:genetics; D018408:Patch-Clamp Techniques; D017434:Protein Structure, Tertiary / Q000235:genetics; D017393:RNA Editing / Q000502:physiology*; D011963:Receptors, GABA-A / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D018682:GABA Agents; C546257:GABRA3 protein, human; D011963:Receptors, GABA-A; D011994:Recombinant Proteins; D007532:Isoleucine; D005680:gamma-Aminobutyric Acid; D008715:Methionine