title: A hot spot for hotfoot mutations in the gene encoding the delta2 glutamate receptor.
abstract: The orphan glutamate receptor delta2 is selectively expressed in Purkinje cells and plays a crucial role in cerebellar functions. Recently, ataxia in the hotfoot mouse ho4J was demonstrated to be caused by a deletion in the delta2 receptor gene (Grid2) removing the N-terminal 170 amino acids of the delta2 receptor. To understand how delta2 receptors function, we characterized mutations in eight additional spontaneously occurring hotfoot alleles of Grid2. The mouse Grid2 gene consists of 16 exons, spanning approximately 1.4 Mb. Genomic DNA analysis showed that seven hotfoot mutants had a deletion of one or more exons encoding the N-terminal domain of delta2 receptors. The exception is ho5J, which has a point mutation in exon 12. Deletions in ho7J, ho9J, ho11J and ho12J mice result in the in-frame deletion of between 40 and 95 amino acids. Expression of constructs containing these deletions in HEK293 cells resulted in protein retention in the endoplasmic reticulum or cis-Golgi without transport to the cell surface. Coimmunoprecipitation assays indicated that these deletions also reduce the intermolecular interaction between individual delta2 receptors. These results indicate that the deleted N-terminal regions are crucial for oligomerization of delta2 receptors and their subsequent transport to the cell surface of Purkinje cells. The relatively large size of the Grid2 gene may be one of the reasons why many spontaneous mutations occur in this gene. In addition, the frequent occurrence of in-frame deletions within the N-terminal domain in hotfoot mutants suggests the importance of this domain in the function of delta2 receptors.
authors.affiliation: Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Shinji
authors.initials: S
authors.lastname: Matsuda
authors.forename: Valerie
authors.initials: V
authors.lastname: Drews
authors.forename: Takashi
authors.initials: T
authors.lastname: Torashima
authors.forename: Miriam H
authors.initials: MH
authors.lastname: Meisler
authors.forename: Michisuke
authors.initials: M
authors.lastname: Yuzaki
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence / Q000235:genetics*; D002478:Cells, Cultured; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000502:physiology; D051379:Mice; D008818:Mice, Neurologic Mutants; D009154:Mutation; D010957:Plasmids; D011487:Protein Conformation; D011689:Purkinje Cells / Q000502:physiology*; D017470:Receptors, Glutamate / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion / Q000235:genetics*; D014162:Transfection
chemical_list: D017470:Receptors, Glutamate; C107913:glutamate receptor delta 2