Figure 7.

Effects of the eif4a3 3′-UTR intron on the level and stability of luc reporter mRNA in wt, Δy14 and Δcpb80 strains. Measurements to analyze the effects on reporter genes containing the eif4a3 3′-UTR intron and controls are shown as follows for each: levels of mRNA in total RNA (left), half-lives of mRNA measured by 4TU RNA pulse–chase (middle), and levels of luciferase enzyme activity/mRNA (right). (A) wt (solid bars) and Δy14 (shaded bars) strains contained the luc reporters shown in Figure S4B. (B) wt (solid bars) and Δcbp80 (hatched bars) strains contained the luc reporters shown in Figure S4B. Measurements were obtained as described in Figure 5. (C) RT–qPCR analysis of eif4a3, erf1, y14, arg-2, eif5, and cox-5 mRNAs purified from wt, Δy14, and Δmago strains by immunopurification of mRNPs with anti-Y14 antibody as described in Materials and Methods. The level of each transcript was normalized to the total amount of purified RNA and then normalized to the level of input transcript. Mock immunopurification from wt without antibody (wt-AB) served as the control. (D) RT–qPCR analyses of luc, eif4a3, and arg-2 mRNAs purified from wt and luc reporter strains by immunopurification of mRNPs with anti-Y14 antibody. mRNA enrichment is shown normalized to the enrichment of eif4a3 in immunopurification from wt cells. See also Figure S5 and Figure S6.