The landscape of translational stall sites in bacteria revealed by monosome and disome profiling

Tomoya Fujita, Takeshi Yokoyama, Mikako Shirouzu, Hideki Taguchi, Takuhiro Ito, Shintaro Iwasaki

Research output: Contribution to journalArticlepeer-review


Ribosome pauses are associated with various cotranslational events and determine the fate of mRNAs and proteins. Thus, the identification of precise pause sites across the transcriptome is desirable; however, the landscape of ribosome pauses in bacteria remains ambiguous. Here, we harness monosome and disome (or collided ribosome) profiling strategies to survey ribosome pause sites in Escherichia coli. Compared to eukaryotes, ribosome collisions in bacteria showed remarkable differences: a low frequency of disomes at stop codons, collisions occurring immediately after 70S assembly on start codons, and shorter queues of ribosomes trailing upstream. The pause sites corresponded with the biochemical validation by integrated nascent chain profiling (iNP) to detect polypeptidyl-tRNA, an elongation intermediate. Moreover, the subset of those sites showed puromycin resistance, presenting slow peptidyl transfer. Among the identified sites, the ribosome pause at Asn586 of ycbZ was validated by biochemical reporter assay, tRNA sequencing (tRNA-seq), and cryo-electron microscopy (cryo-EM) experiments. Our results provide a useful resource for ribosome stalling sites in bacteria.

Original languageEnglish
Pages (from-to)290-302
Number of pages13
Issue number3
Publication statusPublished - 2022 Mar


  • iNP
  • ribosome
  • ribosome pause
  • ribosome profiling
  • translation

ASJC Scopus subject areas

  • Molecular Biology


Dive into the research topics of 'The landscape of translational stall sites in bacteria revealed by monosome and disome profiling'. Together they form a unique fingerprint.

Cite this