005. None of these mRNAs was found among the genes showing the greatest reductions in TE in the inhibitor canagliflozin” mutant versus WT. In fact, four of these 20 mRNAs, all containing long 5UTRs with strong predicted secondary structures, appear to be translated more efficiently on depletion of eIF4G, show ing mean TE4G TEWT ratios of 1. 62 0. 46, 1. 37 0. 23, 1. 24 0. 05, and 1. 24 0. 03. These results do not support the possibility that the translation Inhibitors,Modulators,Libraries of mRNAs with highly stable secondary structures in their 5UTRs would be strongly enhanced by eIF4G. It has been reported that mammalian eIF4G plays a critical role in the ability of post termination 40S subu nits to resume scanning following translation of a short uORF. Hence, we asked whether the genes whose translation is relatively lower in the mutant versus WT might display an atypical occurrence of uORFs.

For the 70 genes with TE4G TEWT values 0. 71 whose occur rences of uORFs were tabulated by Inhibitors,Modulators,Libraries Lawless et al, there is an average of 0. 43 0. 17 uORFs per transcript. For the 47 genes with TE4G TEWT 1. 4, the correspond ing average is 0. 51 0. 26 uORFs per transcript. Neither of these frequencies differs significantly from the average uORF occurrence of 0. 36 0. 02 uORFs Inhibitors,Modulators,Libraries per transcript tabulated for 4149 genes by Lawless et al. Thus, we found no indication that the presence or absence of uORFs is a critical determinant of the effect of eIF4G on the translational efficiency of eIF4G responsive mRNAs. In animals, translational control of specific mRNAs frequently involves Inhibitors,Modulators,Libraries trans acting factors that bind to spe cific recognition elements in the 3UTR and target eIF4F assembly at the cap structure.

Accordingly, we examined whether the 3UTR length differs significantly between the two sets of genes identified above. As shown in Figure 7, the 3UTR length appears to be slightly smaller for the group of genes with TE4G TEWT 0. 71 versus that with TE4G TEWT 1. 4, however, neither group displays a mean 3UTR length that Inhibitors,Modulators,Libraries is significantly different from that of all genes. Hence, it seems unlikely that 3UTR length is an impor tant parameter in determining the dependence of trans lational efficiency on eIF4G. Finally, we examined 10 mRNAs reported to have an A rich IRES and also the IRES containing mRNA URE2, to determine whether the translational efficiencies of these mRNAs might be increased or decreased on depletion of eIF4G.

We observed no significant deviation from unity in the TE4G TEWT ratios of the selleck SB-715992 10 genes with A rich IRESs, ort ORFs require eIF4G to achieve their characteristic, higher than average translational efficiencies Because we examined polysomal RNAs present in heavy polysomes, genes whose transcripts contain, on average, less then 4 translating ribosomes were likely underrepre sented in this analysis.