title: Alternative splicing of exon 14 determines nuclear or cytoplasmic localisation of fmr1 protein isoforms.
abstract: Impaired expression of the FMR1 gene is responsible for the fragile X mental retardation syndrome. The FMR1 gene encodes a cytoplasmic protein with RNA-binding properties. Its complex alternative splicing leads to several isoforms, whose abundance and specific functions in the cell are not known. We have cloned in expression vectors, cDNAs corresponding to several isoforms. Western blot comparison of the pattern of endogenous FMR1 proteins with these transfected isoforms allowed the tentative identification of the major endogenous isoform as ISO 7 and of a minor band as an isoform lacking exon 14 sequences (ISO 6 or ISO 12), while some other isoforms (ISO 4, ISO 5) were not expressed at detectable levels. Surprisingly, in immunofluorescence studies, the transfected splice variants that exclude exon 14 sequences (and have alternate C-terminal regions) were shown to be nuclear. Such differential localisation was however not seen in subcellular fractionation studies. Analysis of various deletion mutants suggests the presence of a cytoplasmic retention domain encoded in exon 14 and of a nuclear association domain encoded within the first eight exons that appear however to lack a typical nuclear localisation signal.
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, INSERM, Strasbourg, France.
authors.forename: A
authors.initials: A
authors.lastname: Sittler
authors.forename: D
authors.initials: D
authors.lastname: Devys
authors.forename: C
authors.initials: C
authors.lastname: Weber
authors.forename: J L
authors.initials: JL
authors.lastname: Mandel
mesh_terms: D016475:3T3 Cells; D017398:Alternative Splicing / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry*; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000737:chemistry*; D005091:Exons / Q000235:genetics*; D051860:Fragile X Messenger Ribonucleoprotein 1; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000032:analysis; D017384:Sequence Deletion
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C497683:FMR1 protein, human; C497684:Fmr1 protein, mouse