title: Polypyrimidine tract-binding protein represses splicing of a fibroblast growth factor receptor-2 gene alternative exon through exon sequences.
abstract: The fibroblast growth factor receptor (FGFR)-2 gene contains two mutually exclusive exons, K-SAM and BEK. We made a cell line designed to become drug-resistant on repression of BEK exon splicing. One drug-resistant derivative of this line carried an insertion within the BEK exon of a sequence containing at least two independent splicing silencers. One silencer was a pyrimidine-rich sequence, which markedly increased binding of polypyrimidine tract-binding protein to the BEK exon. The BEK exon binds to polypyrimidine tract-binding protein even in the silencer's absence. Several exonic pyrimidine runs are required for this binding, and they are also required for overexpression of polypyrimidine tract-binding protein to repress BEK exon splicing. These results show that binding of polypyrimidine tract-binding protein to exon sequences can repress splicing. In epithelial cells, the K-SAM exon is spliced in preference to the BEK exon, whose splicing is repressed. Mutation of the BEK exon pyrimidine runs decreases this repression. If this mutation is combined with the deletion of a sequence in the intron upstream from the BEK exon, a complete switch from K-SAM to BEK exon splicing ensues. Binding of polypyrimidine tract binding protein to the BEK exon thus participates in the K-SAM/BEK alternative splicing choice.
authors.affiliation: INSERM U463, Institut de Biologie-CHR, 9 Quai Moncousu, 44093 Nantes Cedex 1, France.
authors.forename: C
authors.initials: C
authors.lastname: Le Guiner
authors.forename: A
authors.initials: A
authors.lastname: Plet
authors.forename: D
authors.initials: D
authors.lastname: Galiana
authors.forename: M C
authors.initials: MC
authors.lastname: Gesnel
authors.forename: F
authors.initials: F
authors.lastname: Del Gatto-Konczak
authors.forename: R
authors.initials: R
authors.lastname: Breathnach
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D004247:DNA; D005091:Exons; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D038941:Polypyrimidine Tract-Binding Protein; D016601:RNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics*; D012261:Ribonucleoproteins / Q000378:metabolism / Q000502:physiology*
chemical_list: D016601:RNA-Binding Proteins; D017468:Receptors, Fibroblast Growth Factor; D012261:Ribonucleoproteins; D038941:Polypyrimidine Tract-Binding Protein; D004247:DNA; C496353:FGFR2 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051497:Receptor, Fibroblast Growth Factor, Type 2