A peptide motif in Raver1 mediates splicing repression by interaction with the PTB RRM2 domain.
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.
Rideau
Alexis P
AP
Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, CB2 1GA, UK.
Gooding
Clare
C
Simpson
Peter J
PJ
Monie
Tom P
TP
Lorenz
Mike
M
Hüttelmaier
Stefan
S
Singer
Robert H
RH
Matthews
Stephen
S
Curry
Stephen
S
Smith
Christopher W J
CW
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
D002352:Carrier Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D038941:Polypyrimidine Tract-Binding Protein; D012313:RNA; D011392:Proline