The WW domain-containing proteins interact with the early spliceosome and participate in pre-mRNA splicing in vivo.
A growing body of evidence supports the coordination of mRNA synthesis and its subsequent processing events. Nuclear proteins harboring both WW and FF protein interaction modules bind to splicing factors as well as RNA polymerase II and may serve to link transcription with splicing. To understand how WW domains coordinate ... the assembly of splicing complexes, we used glutathione S-transferase fusions containing WW domains from CA150 or FBP11 in pull-down experiments with HeLa cell nuclear extract. The WW domains associate preferentially with the U2 small nuclear ribonucleoprotein and with splicing factors SF1, U2AF, and components of the SF3 complex. Accordingly, WW domain-associating factors bind to the 3' part of a pre-mRNA to form a pre-spliceosome-like complex. We performed both in vitro and in vivo splicing assays to explore the role of WW/FF domain-containing proteins in this process. However, although CA150 is associated with the spliceosome, it appears to be dispensable for splicing in vitro. Nevertheless, in vivo depletion of CA150 substantially reduced splicing efficiency of a reporter pre-mRNA. Moreover, overexpression of CA150 fragments containing both WW and FF domains activated splicing and modulated alternative exon selection, probably by facilitating 3' splice site recognition. Our results suggest an essential role of WW/FF domain-containing factors in pre-mRNA splicing that likely occurs in concert with transcription in vivo.
Mesh Terms:
Animals, Carrier Proteins, DNA-Binding Proteins, Exons, Hela Cells, Humans, Macromolecular Substances, Mice, Nuclear Proteins, Protein Structure, Tertiary, RNA Polymerase II, RNA Precursors, RNA Splicing, RNA, Messenger, Recombinant Fusion Proteins, Ribonucleoprotein, U2 Small Nuclear, Ribonucleoproteins, Spliceosomes, Trans-Activators, Transcription Factors, Transcription, Genetic
Animals, Carrier Proteins, DNA-Binding Proteins, Exons, Hela Cells, Humans, Macromolecular Substances, Mice, Nuclear Proteins, Protein Structure, Tertiary, RNA Polymerase II, RNA Precursors, RNA Splicing, RNA, Messenger, Recombinant Fusion Proteins, Ribonucleoprotein, U2 Small Nuclear, Ribonucleoproteins, Spliceosomes, Trans-Activators, Transcription Factors, Transcription, Genetic
Mol. Cell. Biol.
Date: Oct. 01, 2004
PubMed ID: 15456888
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