Effects of cycloheximide on the interpretation of ribosome profiling experiments in Schizosaccharomyces pombe
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Duncan, D., & Mata, J. (2017). Effects of cycloheximide on the interpretation of ribosome profiling experiments in Schizosaccharomyces pombe. Scientific Reports, 7 (10331)https://doi.org/10.1038/s41598-017-10650-1
Stress conditions lead to global and gene-specific changes in RNA translation. Ribosome profiling experiments have identified genome-wide alterations in the distribution of ribosomes along mRNAs. However, it is contentious whether these changes reflect real responses, or whether they are artefacts caused by the use of inhibitors of translation (notably cycloheximide). To address this issue we performed ribosome profiling with the fission yeast Schizosaccharomyces pombe under conditions of exponential growth (unstressed) and nitrogen starvation (nutritional stress), and both in the presence and absence of cycloheximide. We examined several aspects of the translational response, including density of ribosomal footprints on coding sequences, 5’ leader ribosomal densities, distribution of ribosomes along coding sequences, and ribosome codon occupancies. Cycloheximide had minor effects on overall ribosome density, which affected mostly mRNAs encoding ribosomal proteins. Nitrogen starvation caused an accumulation of ribosomes on 5’ leaders in both cycloheximide-treated and untreated cells. By contrast, stress-induced ribosome accumulation on the 5’ side of coding sequences was cycloheximide-dependent. Finally, codon occupancy showed strong positive correlations in cycloheximide-treated and untreated cells. Our results demonstrate that cycloheximide does influence some of the results of ribosome profiling experiments, although it is not clear if this effect is always artefactual.
This work was supported by a grant from the Biotechnology and Biological Sciences Research Council (BBSRC) [BB/M021483/1].
External DOI: https://doi.org/10.1038/s41598-017-10650-1
This record's URL: https://www.repository.cam.ac.uk/handle/1810/267506
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