Unphosphorylated SR-Like Protein Npl3 Stimulates RNA Polymerase II Elongation
Figure 5
Opposing effects in termination demonstrated using RNA-binding or phosphorylation defective npl3 alleles.
RNA was extracted from NPL3, npl3-120 and npl3-S411A and whole transcript sense strand cDNA was synthesized and used for hybridization to tiled arrays. The hybridization signals for (A) FBA1, MPE1 and (B) TDH3 for two independent experiments were used to calculate the ratio of mutant vs. wild-type. The top panel shows the ratio of rna15-2/RNA15; bottom panel shows the ratio of each npl3 mutant/NPL3; a solid line for npl3-120/NPL3, or dashed line for npl3-S411A/NPL3. The corresponding position and orientation (Watson (W+), dashed bars, or Crick (C-) strand, black solid bars) of the genes is also shown. (C–D) A 300 bp region at the beginning of each ORF (+50 to +350 relative to the start codon), was selected for comparison to the same size region at the 3′UTR (−100 to +200 relative to the stop codon) of the genes that showed increase readthrough for the npl3-S411A strain. The mean value of the log2 ratio of npl3-S411A/NPL3 was calculated and graphed in a scatter plot. (D) The total number of genes corresponding to the Crick strand were used to calculate the mean value of the same region described for (C).