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. Author manuscript; available in PMC: 2019 Apr 19.
Published in final edited form as: Mol Cell. 2018 Apr 19;70(2):254–264.e6. doi: 10.1016/j.molcel.2018.03.015

Figure 3. Translation elongation pause enhances initiation at upstream AUU codon.

Figure 3

(A) Amino acid sequence of the mouse azin1 uCC. The last 10 residues (red) were replaced by (bottom) the C-terminal two-thirds of the N. crassa AAP sequence (green) in uCC-AAP hybrid constructs; critical APP residue D12 is boxed in orange. (B) The indicated uCC-Luc reporters or a dual-luciferase human OAZ1 frameshift reporter (to monitor polyamine levels) were transfected either in HEK293T cells (right panel) pretreated with DFMO or DFMO + 3 mM SPD or in U2OS cells (middle panel) cultured in DMEM (400 μM Arg) media +/- 25 mM Arg. Percent AUU initiation calculation and firefly normalization were performed as in Figure 1. Error bars denote standard deviation; *p<0.05 (Student’s two-tailed t-test; n=4, assayed in duplicate); relative reporter mRNA levels are presented in Table S2.