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. 2010 Oct;16(10):2033–2041. doi: 10.1261/rna.2366310

FIGURE 3.

FIGURE 3.

Deletion of CBP80 is synthetic with L30 repression of splicing. (A) Decreased levels of Cbp80-TAP in SLR5 mutants. Cbp80 protein was TAP-tagged at the C terminus in yJV25 (wt, lane 1), SLR5 (lane 2), and cbp20Δ (Y02074). Extracts were subjected to Western analyses, as indicated. Tubulin was used as loading control. (B) Deletion of CPB80 produces the same phenotype as that of SLR5. One-fifth serial dilutions of wt (yJV25, top), cbp80Δ (yJV35, middle), and SLR5 (bottom) cells transformed with pLCUP 5A C9T were spotted on copper-containing media, as indicated. Copper sensitivity indicates splicing repression of the LCUP 5A C9U transcript (C). Deletion of CBP80 leads to repression of LCUP 5A C9U splicing by L30. Northern analysis of RNA from wt (yJV25, lane 1), SLR5 (lane 2), and cbp80Δ (lane 3) cells. Positions of precursor (p) and mature (m) LGFP 5A C9U are indicated on the right. (D) Splicing efficiency and repression by L30 of the RPL30 intron in wt and cbp80Δ cells, with or without excess L30. Northern analysis of RNA from either wt (even lanes) or cbp80Δ cells (odd lanes) transformed with the pLGFP reporter plasmid, as indicated at the top. Samples in the bottom panel are from cells under excess L30 (pMB73). Positions of precursor (p) and mature (m) LGFP are indicated on the left. pLGFP contains the GFP ORF instead of Cup1 in pLCUP (Vilardell and Warner 1997). pMB73 encodes L30 without the autoregulatory loop (Macías et al. 2008).