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. 2010 Aug 1;24(15):1645–1658. doi: 10.1101/gad.1940010

Figure 6.

Figure 6.

A Ur element RNA oligonucleotide alters SL trans-splicing specificity. (A, top) Diagram of wild-type rla-1 substrate, as in Figure 1. Numbers 1, 2, and 3 indicate sites of SL2 trans-splicing in the presence of a Ur element RNA oligo. (Middle) Diagram of long RNA oligos added to in vitro splicing reactions. The long Ur element oligo (Ur) includes the stem and all of the UAYYUU motifs (each indicated by a horizontal bar above the sequence). The control RNA oligo (C) contains substitutions, indicated by shaded boxes. (Bottom) SL1, SL2, and exon RT–PCR of in vitro splicing reactions of the wild-type rps-3 and rla-1 RNA substrates (shown in Fig. 1 and in the top panel) in the absence (−) or presence of an excess amount of control or long Ur element RNA oligo. Percent is calculated relative to the no oligo lanes. (B, left) Diagrams of short RNA oligos added to in vitro splicing reactions. The wild-type short Ur element sequence contains the stem–loop and one UAYYUU motif (horizontal bar above the sequence). Substitutions in other oligos are indicated by shaded boxes. (Right) SL2 RT–PCR of in vitro splicing reactions of the wild-type rla-1 substrate in the absence (−) or presence of indicated RNA oligos. Spliced products 1, 2, and 3 are SL2 trans-spliced at the sites indicated in the diagram in A.