title: Proline-rich sequence recognition: I. Marking GYF and WW domain assembly sites in early spliceosomal complexes.
abstract: Proline-rich sequences (PRS) and their recognition domains have emerged as transposable protein interaction modules during eukaryotic evolution. They are especially abundant in proteins associated with pre-mRNA splicing and likely assist in the formation of the spliceosome by binding to GYF and WW domains. Here we profile PRS-mediated interactions of the CD2BP2/52K GYF domain by a site-specific peptide inhibitor and stable isotope labeling/mass spectrometry analysis. Several PRS hubs with multiple proline-rich motifs exist that can recruit GYF and/or WW domains. Saturating the PRS sites by an isolated GYF domain inhibited splicing at the level of A complex formation. The interactions mediated by PRS are therefore important to the early phases of spliceosomal assembly.
authors.affiliation: Protein Engineering Group, Leibniz Institute for Molecular Pharmacology and Freie Universität Berlin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Kofler
authors.forename: Michael
authors.initials: M
authors.lastname: Schuemann
authors.forename: Christian
authors.initials: C
authors.lastname: Merz
authors.forename: Daniela
authors.initials: D
authors.lastname: Kosslick
authors.forename: Andreas
authors.initials: A
authors.lastname: Schlundt
authors.forename: Astrid
authors.initials: A
authors.lastname: Tannert
authors.forename: Michael
authors.initials: M
authors.lastname: Schaefer
authors.forename: Reinhard
authors.initials: R
authors.lastname: Lührmann
authors.forename: Eberhard
authors.initials: E
authors.lastname: Krause
authors.forename: Christian
authors.initials: C
authors.lastname: Freund
mesh_terms: D001665:Binding Sites; D002384:Catalysis; D002460:Cell Line; D003001:Cloning, Molecular; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods; D010455:Peptides / Q000737:chemistry; D011392:Proline / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism*
chemical_list: D010455:Peptides; D012333:RNA, Messenger; D011392:Proline