title: Ultrastructural analysis of the functional domains in FMRP using primary hippocampal mouse neurons.
abstract: Fragile X syndrome is caused by lack of the protein FMRP. FMRP mediates mRNA binding, dendritic mRNA transport and translational control at spines. We examined the role of functional domains of FMRP in neuronal RNA-granule formation and dendritic transport using different FMRP variants, including the mutant FMRP_I304N and the splice-variant FMRP_Iso12. Both variants are absent from dendritic RNA-granules in Fmr1 knockout neurons. Co-transfection experiments showed that wild-type FMRP recruits both FMRP variants into dendritic RNA-granules. Co-transfection of FXR2, an FMRP homologue, also resulted in redistribution of both variants into dendritic RNA-granules. Furthermore, the capacity of the variants to transport their mRNAs and the mRNA localization of an FMR1 construct containing silent point-mutations affecting only the G-quartet-structure were investigated. In conclusion, we show that wild-type FMRP and FXR2P are able to recruit FMRP variants into RNA-granules and that the G-quartet-structure in FMR1 mRNA is not essential for its incorporation in RNA-granules.
authors.affiliation: CBG Department of Clinical Genetics, Erasmus MC, Dr. Molewaterplein 50, 3015 GE, P.O. Box 2040, Rotterdam 3000 CA, The Netherlands.
authors.forename: Josien
authors.initials: J
authors.lastname: Levenga
authors.forename: Ronald A M
authors.initials: RA
authors.lastname: Buijsen
authors.forename: Maria
authors.initials: M
authors.lastname: Rifé
authors.forename: Hervé
authors.initials: H
authors.lastname: Moine
authors.forename: David L
authors.initials: DL
authors.lastname: Nelson
authors.forename: Ben A
authors.initials: BA
authors.lastname: Oostra
authors.forename: Rob
authors.initials: R
authors.lastname: Willemsen
authors.forename: Femke M S
authors.initials: FM
authors.lastname: de Vrij
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003712:Dendrites / Q000378:metabolism* / Q000648:ultrastructure; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D034443:RNA Transport; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; D020033:Protein Isoforms; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C512309:FXR2 protein, mouse; C497684:Fmr1 protein, mouse