Proline-rich sequence recognition: I. Marking GYF and WW domain assembly sites in early spliceosomal complexes.
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.
Kofler
Michael
M
Protein Engineering Group, Leibniz Institute for Molecular Pharmacology and Freie Universität Berlin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
Schuemann
Michael
M
Merz
Christian
C
Kosslick
Daniela
D
Schlundt
Andreas
A
Tannert
Astrid
A
Schaefer
Michael
M
Lührmann
Reinhard
R
Krause
Eberhard
E
Freund
Christian
C
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*
D010455:Peptides; D012333:RNA, Messenger; D011392:Proline