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. Author manuscript; available in PMC: 2009 Apr 14.
Published in final edited form as: Nat Struct Mol Biol. 2008 Sep 14;15(10):1040–1048. doi: 10.1038/nsmb.1485

Figure 5.

Figure 5

SRp38 binds to and activates splicing of RNA substrates containing the GluR-B Flip or Flop exon. (a) Comparison of a putative SRp38 binding motif in Flip and Flop exons from the murine GluR-B transcript with the SRp38 consensus recognition sequence. (b) Flop and Flip exons of the GluR-B pre-mRNA bind SRp38 specifically. Gel-shift assays with the indicated radiolabeled RNAs were performed with increasing amounts of GST-SRp38 (25 ng, 75 ng and 225 ng). Complexes were resolved by nondenaturing PAGE. (c) Flop and Flip exons of the GluR-B pre-mRNA do not bind His-SC35. Increasing amounts of purified His-SC35 (25 ng, 75 ng and 225 ng) were added to gel-shift assays containing the identical RNAs. (d) SRp38 activates splicing of RNA substrates containing Flip or Flop exons. The downstream β-globin exon was replaced with Flip or Flop exons, and splicing reactions were performed in S100 with increasing amounts of GST-SRp38 (20 ng, 40 ng and 100 ng) in the presence or absence of NF40–60 as indicated.