Translation elongation factor 4 (LepA) contributes to tetracycline susceptibility by stalling elongating ribosomes

Purpose: We use the ribosome profiling protocol to understand EF4 mediated translation events. Methods: We used ribosome profiling data to analyze by plastid software. The ribosome were purified by sucrose gradient separation. Results: Using sequencing data, we found that EF4 mediates the 1-nucleotide conformational change of ribosomes. We also found that many genes involved in translation after tetracycline treatment. Finally, we found that EF4 stalls the elongating ribosomes globally. Conclusions: Our results suggest that EF4 mediates both 30S biogenesis and translation elongation process, in particular, EF4 stalls the elongating ribosomes globally. Overall design: Ribosome footprints from WT and ?EF4 strains and the two strains treated with tetracycline were sequenced, in two replicates, using Illumina HiSeq 2500.