1. Chromosomes and Gene Expression
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Translation: Finding sense in the context

  1. Kim M Keeling
  2. David M Bedwell  Is a corresponding author
  1. Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, United States
  2. Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, United States
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Cite this article as: eLife 2020;9:e55960 doi: 10.7554/eLife.55960
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Using ribosomal profiling to assess the efficiency of translation termination.

At any given time, a number of ribosomes (grey ovals) will be attached to messenger RNA molecules at different positions. Ribosomal profiling is a technique that can help to locate ribosomes on the mRNA they are translating at the scale of the genome. First, each ribosome (and the region of mRNA it is attached to) is isolated. These segments are then treated with nucleases, releasing the RNA sequences the ribosomes were bound to. These are then purified and sequenced. During efficient termination (left), most of the ribosomes are between the start codon (green) and the stop codon (red), with the majority accumulating at the stop codon (graph on the bottom left), where translation stops. During inefficient termination (right), for example under the influence of small molecules such as G418, ribosomes can ‘readthrough’ the termination codon and carry on translating downstream (graph on the bottom right).

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