Mechanisms involved in the reduction of GABAA receptor alpha1-subunit expression caused by the epilepsy mutation A322D in the trafficking-competent receptor.
A mutation in the alpha1-subunit (A322D) of GABA(A)Rs is responsible for juvenile myoclonic epilepsy in a large Canadian family. Previous work has identified that this mutant affects the cell expression and function of recombinant GABA(A)Rs, expressed in HEK293 cells. Here we have extended these observations by showing that the mutation promotes association with the endoplasmic reticulum chaperone calnexin and accelerates the degradation rate of the subunits approximately 2.5-fold. We also find that the mutation causes the subunit to be degraded largely by a lysosomal-dependent process. Furthermore, we find that the mutation results in receptors that are inserted into the plasma membrane but are more rapidly endocytosed by a dynamin and caveolin1-dependent mechanism. These results suggest that the mutant subunit can form functional receptors, but that these have a shorter lifetime on the plasma membrane.
Brain Research Centre, University of British Columbia, Vancouver V6T 1Z3, Canada.
Clarrisa A
CA
Bradley
Changiz
C
Taghibiglou
Graham L
GL
Collingridge
Yu Tian
YT
Wang
D037281:Calnexin / Q000378:metabolism; D002460:Cell Line; D004705:Endocytosis; D005786:Gene Expression Regulation; D006801:Humans; D009154:Mutation; D020190:Myoclonic Epilepsy, Juvenile / Q000235:genetics*; D011963:Receptors, GABA-A / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
C546240:GABRA1 protein, human; D011963:Receptors, GABA-A; D011994:Recombinant Proteins; D037281:Calnexin