Ribosome profiling uncovers selective mRNA translation associated with eIF2 phosphorylation in erythroid progenitors

The aim of this study is to investigate how eIF2 phosphorylation affects mRNA translation in erythroblasts. Ribosome profiling combined with RNA sequencing was used to determine translation initiation sites and ribosome density on individual transcripts. Ribosome profiling samples with translation halted by harringtonine (ribosome stalls at start site) or cyclohexamide (ribosome is frozen at current location) were treated with tunicamycin to induce eIF2 phosphorylation vs untreated. Total RNAseq sample with and without tunicamcyin were also generated. Total RNAseq and cycolhexamide were used to calculate ribosome occupancy as a measure of translation efficiency. Harringtonin peaks were used to identify alternative sites of translation initiation under stress.