Ribosome profiling of the genome-reduced bacterium Mycoplasma pneumoniae

In order to study quantitatively the translation efficiency in the genome-reduced bacterium Mycoplasma pneumoniae, we performed ribosome profiling (ribo-seq) in standard growth conditions. We first tested whether the ribosome profiling method could be applied to M. pneumoniae and assessed the quality of the obtained data. Mpn present some characteristics that differ from other model bacterial species where ribosome profiling has been established, like the absence of a cell wall, different lipid composition and low number of ribosomes (200-300 per cell compared with ~55,000 in E. coli). Thus, several aspects of the original method (Ingolia et al., 2009) had to be adapted. As a reference, we also performed ribosome profiling in E. coli. We also calibrated the ribosome P-site shift from the 3' end of ribosome protected footprints (RPFs) by analyzing the metagene profile of a sample treated with tetracycline.