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. Author manuscript; available in PMC: 2016 Jul 2.
Published in final edited form as: Cell. 2015 Jun 18;162(1):59–71. doi: 10.1016/j.cell.2015.05.049

Figure 5. The 5′UTR confers translational sensitivity to eIF4E target mRNAs.

Figure 5

A) Diagram of the 5′UTR luciferase reporter assay. B) Requirements for eIF4E in 5′UTR mediated translation of target mRNAs (Fth1, Lmnb1, Gclc, Edn1, and Pdgfrb) and control mRNAs (Gapdh and B2M) by 5′UTR luciferase reporter assay in transformed MEFs. “pGL3” denotes the test vector lacking a 5′UTR. Results are normalized to 5′UTR reporter activity in transformed WT cells. C) Comparison of canonical 5′UTR features between all mouse genes and the subset whose translation efficiency is reduced in Eif4e+/− during oncogenic transformation. D) Consensus sequence and enrichment value (E-value) of the Cytosine Enriched Regulator of Translation (CERT) motif identified by MEME analysis along with a diagram illustrating the frequency of eIF4E target mRNAs containing a CERT. E) Effect of C to G transversion mutations of the CERT domains in Fth1 (+153 nt from 5′cap and −1nt from the ATG) and in Edn1 (+422 nt from the 5′cap and −188nt from the ATG) on 5′UTR luciferase reporter activity in transformed WT and Eif4e+/− MEFs. Results are normalized to non-mutated 5′UTR reporter activity in transformed WT cells. Asterisks indicate a statistically significant change (*= p<0.05, **= p<0.01). “n.s.” = not significant. All values represent the mean +SEM except for (C) which represent mean +SD.