title: Structural basis for the recognition of spliceosomal SmN/B/B' proteins by the RBM5 OCRE domain in splicing regulation.
abstract: The multi-domain splicing factor RBM5 regulates the balance between antagonistic isoforms of the apoptosis-control genesFAS/CD95,Caspase-2andAID. An OCRE (OCtamer REpeat of aromatic residues) domain found in RBM5 is important for alternative splicing regulation and mediates interactions with components of the U4/U6.U5 tri-snRNP. We show that the RBM5 OCRE domain adopts a unique β-sheet fold. NMR and biochemical experiments demonstrate that the OCRE domain directly binds to the proline-rich C-terminal tail of the essential snRNP core proteins SmN/B/B'. The NMR structure of an OCRE-SmN peptide complex reveals a specific recognition of poly-proline helical motifs in SmN/B/B'. Mutation of conserved aromatic residues impairs binding to the Sm proteinsin vitroand compromises RBM5-mediated alternative splicing regulation of FAS/CD95. Thus, RBM5 OCRE represents a poly-proline recognition domain that mediates critical interactions with the C-terminal tail of the spliceosomal SmN/B/B' proteins inFAS/CD95alternative splicing regulation.
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München, Neuherberg, Germany.|Biomolecular NMR and Center for Integrated Protein Science Munich, Department Chemie, Technische Universität München, Garching, Germany.
authors.forename: André
authors.initials: A
authors.lastname: Mourão
authors.affiliation: Barcelona Institute of Science and Technology and Universitat Pompeu Fabra, Centre de Regulació Genòmica, Barcelona, Spain.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bonnal
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München, Neuherberg, Germany.|Biomolecular NMR and Center for Integrated Protein Science Munich, Department Chemie, Technische Universität München, Garching, Germany.
authors.forename: Komal
authors.initials: K
authors.lastname: Soni
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München, Neuherberg, Germany.|Biomolecular NMR and Center for Integrated Protein Science Munich, Department Chemie, Technische Universität München, Garching, Germany.
authors.forename: Lisa
authors.initials: L
authors.lastname: Warner
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS-UMR5535, Université de Montpellier, Montpellier, France.
authors.forename: Rémy
authors.initials: R
authors.lastname: Bordonné
authors.affiliation: Barcelona Institute of Science and Technology and Universitat Pompeu Fabra, Centre de Regulació Genòmica, Barcelona, Spain.|Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Valcárcel
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München, Neuherberg, Germany.|Biomolecular NMR and Center for Integrated Protein Science Munich, Department Chemie, Technische Universität München, Garching, Germany.
authors.forename: Michael
authors.identifier: http://orcid.org/0000-0002-1594-0527
authors.initials: M
authors.lastname: Sattler
mesh_terms: D019943:Amino Acid Substitution; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D009682:Magnetic Resonance Spectroscopy; D011392:Proline / Q000378:metabolism; D011485:Protein Binding; D000072757:Protein Conformation, beta-Strand; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016823:Volvocida / Q000201:enzymology / Q000378:metabolism; D019014:fas Receptor / Q000378:metabolism; D055517:snRNP Core Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: C491901:FAS protein, human; D016601:RNA-Binding Proteins; D019014:fas Receptor; D055517:snRNP Core Proteins; D011392:Proline
keywords: NMR-spectroscopy; OCRE domain; alternative splicing; biochemistry; biophysics; human; poly-proline; protein-protein interactions; structural biology