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A systematically-revised ribosome profiling method for bacteria reveals pauses at single-codon resolution
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A systematically-revised ribosome profiling method for bacteria reveals pauses at single-codon resolution
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Journal Article
A systematically-revised ribosome profiling method for bacteria reveals pauses at single-codon resolution
2019
Overview
In eukaryotes, ribosome profiling provides insight into the mechanism of protein synthesis at the codon level. In bacteria, however, the method has been more problematic and no consensus has emerged for how to best prepare profiling samples. Here, we identify the sources of these problems and describe new solutions for arresting translation and harvesting cells in order to overcome them. These improvements remove confounding artifacts and improve the resolution to allow analyses of ribosome behavior at the codon level. With a clearer view of the translational landscape in vivo, we observe that filtering cultures leads to translational pauses at serine and glycine codons through the reduction of tRNA aminoacylation levels. This observation illustrates how bacterial ribosome profiling studies can yield insight into the mechanism of protein synthesis at the codon level and how these mechanisms are regulated in response to changes in the physiology of the cell.
Publisher
eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
