Figure 7: Prediction and validation of conserved uORFs.
From: Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae
Conserved uORFs show a 5–10 times repressive effect on reporter gene translation. a, Alignments of two tested uORFs. uORFs are shown in purple boxes, and protein-coding genes in a blue background. Conserved bases are in upper case and start/stop codons are highlighted. b, Experimental design. The A. nidulans sequence from 26 nucleotides upstream of each uORF to four codons in the protein-coding gene was fused with a firefly luciferase gene. Controls were generated with start codons for both the uORF and the luciferase gene (+/+), the luciferase gene only (-/+), and the uORF only (+/-). Starts were deleted by alteration to ATT. mRNA from each construct was used to programme a cell-free translation system. c, Results of translation assays. The luciferase activity of all constructs (normalized to the -/+ construct) is shown on the y axis. Error bars show the average of the absolute deviation from the mean. The autoradiogram shows 35S-Met-labelled firefly luciferase obtained by in vitro translation of the same mRNAs.
