Development of a tissue-specific ribosome profiling approach in Drosophila enables genome-wide evaluation of translational adaptations
Fig 5
Analysis of transcriptional and translational profiling of GluRIIA expression in GluRIIA mutants and Tor overexpression in Tor-OE.
(A) Schematic illustrating the genomic GluRIIA locus in wild type and GluRIIASP16 mutants. Note that the 5’ region of GluRIIA is deleted in the GluRIIASP16 mutant, as well as the adjacent oscillin gene. Below: RNA-seq reads mapping to the GluRIIA locus from transcriptional and ribosome profiling in wild type and GluRIIASP16 mutants. The coverage graphs were divided into four sections corresponding to the regions indicated in the GluRIIA transcript. The numbers in each graph indicates the expression value of that region normalized to wild type transcriptional or ribosome profiling expression value. Note that no expression was detected by transcriptional or ribosome profiling in the deleted region in GluRIIA mutants, as expected. (B) Schematic illustrating the endogenous Tor mRNA transcript and the mRNA transcript transgenically expressed in Tor-OE (UAS-Tor). Both transcripts share the same coding sequence, but differ in their 5’UTR and 3’UTR sequences. Below are reads mapping to the indicated regions, divided into the three indicated sections. Note that both transcriptional and translational expression of UAS-Tor mRNA are significantly increased in Tor-OE, while transcription and translation of endogenous Tor mRNA is largely unchanged in Tor-OE. (C) Histogram of the distribution of gene translation changes in wild type versus wild type (black), which represents intrinsic variability, that of Tor-OE versus wild type (Red), and that of GluRIIA mutants versus wild type (blue). Note the shift in distribution observed in Tor-OE, suggesting a global increase in translation. (D) Cumulative percentage plot of distributions shown in (C), showing significant difference between Tor-OE versus wild type distribution compared to wild type versus wild type distribution. (p<0.001, Kolmogorov–Smirnov test).