Fig. 9

mRNAs that show reciprocal changes in the translation of uORFs and mORFs at multiple temperatures. a Wiggle track images showing ribosome-protected fragments (RPF) on the GCN4 mRNA in cells cultured at either 20, 30, or 37 °C, in units of rpm (reads per million mapped reads from two replicates at each temperature). The RPF tracks were normalized to the mRNA levels at each temperature to reflect the changes in translation efficiencies (ΔTE) of uORF and mORF as described in the legend to Fig. 2 and the “Materials and methods” section. The schematic shows the position of the uORFs (purple rectangles) and mORF (striped pink rectangle). NCC uORFs are shown with striped purple rectangle. AUG uORFs are in purple rectangles. Average change in the TEs of the three NCC uORFs showing significant changes in translation at 20 or 37 °C (Avg. ∆TE_{NCC uORFs}) is shown. The enlargement of the boxed area is also shown below with start sites of NCC uORFs (bold, underlined) and the − 3 to − 1 and + 4 context nucleotides. The green arrow shows the NCC uORF start site (AUA) that has been previously shown to be used as an upstream start site [46]. b–g Same as in a but for b the CPA1 mRNA, c ADH4 mRNA, d Dur1,2 mRNA, e ATG40 mRNA, f AGA1 mRNA, g AGA2 mRNA. h, i Wiggle track images of the ALA1 (h) and GRS1 (i) mRNAs as described in a. The N-terminal extension (striped purple rectangle) and mORF (striped pink rectangle) are shown. Relative TE_NTD (TE_NTE/TE_mORF ratio) reflects the ratio of translation efficiency of initiation at the start site of the NTE (ACG in the case of ALA1 and UUG in the case of GRS1) to that of the combined initiation events at NTE and mAUG