title: A peptide motif in Raver1 mediates splicing repression by interaction with the PTB RRM2 domain.
abstract: Polypyrimidine tract-binding protein (PTB) is a regulatory splicing repressor. Raver1 acts as a PTB corepressor for splicing of alpha-tropomyosin (Tpm1) exon 3. Here we define a minimal region of Raver1 that acts as a repressor domain when recruited to RNA. A conserved [S/G][I/L]LGxxP motif is essential for splicing repressor activity and sufficient for interaction with PTB. An adjacent proline-rich region is also essential for repressor activity but not for PTB interaction. NMR analysis shows that LLGxxP peptides interact with a hydrophobic groove on the dorsal surface of the RRM2 domain of PTB, which constitutes part of the minimal repressor region of PTB. The requirement for the PTB-Raver1 interaction that we have characterized may serve to bring the additional repressor regions of both proteins into a configuration that allows them to synergistically effect exon skipping.
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, CB2 1GA, UK.
authors.forename: Alexis P
authors.initials: AP
authors.lastname: Rideau
authors.forename: Clare
authors.initials: C
authors.lastname: Gooding
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Simpson
authors.forename: Tom P
authors.initials: TP
authors.lastname: Monie
authors.forename: Mike
authors.initials: M
authors.lastname: Lorenz
authors.forename: Stefan
authors.initials: S
authors.lastname: Hüttelmaier
authors.forename: Robert H
authors.initials: RH
authors.lastname: Singer
authors.forename: Stephen
authors.initials: S
authors.lastname: Matthews
authors.forename: Stephen
authors.initials: S
authors.lastname: Curry
authors.forename: Christopher W J
authors.initials: CW
authors.lastname: Smith
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D038941:Polypyrimidine Tract-Binding Protein / Q000737:chemistry* / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D038941:Polypyrimidine Tract-Binding Protein; D012313:RNA; D011392:Proline