Gerashchenko et al. 2014 (PRJNA255669)

General Details

Title Translation Inhibitors Cause Abnormalities in Ribosome Profiling Experiments
Organism Saccharomyces cerevisiae
Number of Samples 24
Release Date 2014/07/18 00:00
Sequencing Types
Protocol Details

Study Links

Repository Details

SRA SRP044649
ENA SRP044649
GEO
BioProject PRJNA255669

Publication

Title
Authors Gerashchenko MV,Gladyshev VN
Journal Nucleic acids research
Publication Date 2014
Abstract Ribosome profiling and high-throughput sequencing provide unprecedented opportunities for the analysis of mRNA translation. Using this novel method, several studies have demonstrated the widespread role of short upstream reading frames in translational control as well as slower elongation at the beginning of open reading frames in response to stress. Based on the initial studies, the importance of adding or omitting translation inhibitors, such as cycloheximide, was noted as it markedly affected ribosome coverage profiles. For that reason, many recent studies omitted translation inhibitors in the culture medium. Here, we investigate the influence of ranging cycloheximide concentrations on ribosome profiles in Saccharomyces cerevisiae and demonstrate that increasing the drug concentration can overcome some of the artifacts. We subjected cells to various manipulations and show that neither oxidative stress nor heat shock nor amino acid starvation affect translation elongation. Instead, the observations in the initial studies are the result of cycloheximide-inflicted artifacts. Likewise, we find little support for short upstream reading frames to be involved in widespread protein synthesis regulation under stress conditions. Our study highlights the need for better standardization of ribosome profiling methods. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.
PMC PMC4176156
PMID 25056308
DOI
Run Accession Study Accession Scientific Name Cell Line Library Type Treatment GWIPS-viz Trips-Viz RiboCrypt Reads BAM BigWig (F) BigWig (R)
SRR1520311 PRJNA255669 Saccharomyces cerevisiae BY4741 Frozen
SRR1520312 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520313 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520314 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520315 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520316 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520317 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520318 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520319 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520320 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520321 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520322 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520323 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520324 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520325 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520326 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520327 PRJNA255669 Saccharomyces cerevisiae BY4741 Frozen
SRR1520328 PRJNA255669 Saccharomyces cerevisiae BY4741 Cycloheximide
SRR1520329 PRJNA255669 Saccharomyces cerevisiae BY4741
SRR1520330 PRJNA255669 Saccharomyces cerevisiae BY4741
SRR1520331 PRJNA255669 Saccharomyces cerevisiae BY4741
SRR1520332 PRJNA255669 Saccharomyces cerevisiae BY4741 Frozen
SRR1520333 PRJNA255669 Saccharomyces cerevisiae BY4741 Frozen
SRR1520334 PRJNA255669 Saccharomyces cerevisiae
Run Accession Study Accession Scientific Name Cell Line Library Type Treatment GWIPS-viz Trips-Viz RiboCrypt Reads BAM BigWig (F) BigWig (R)

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