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-Structure paper
タイトル | The landscape of translational stall sites in bacteria revealed by monosome and disome profiling. |
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ジャーナル・号・ページ | RNA, Vol. 28, Issue 3, Page 290-302, Year 2022 |
掲載日 | 2021年12月14日 |
![]() | Tomoya Fujita / Takeshi Yokoyama / Mikako Shirouzu / Hideki Taguchi / Takuhiro Ito / Shintaro Iwasaki / ![]() |
PubMed 要旨 | 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; ...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 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 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. |
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手法 | EM (単粒子) |
解像度 | 3.3 Å |
構造データ | EMDB-30431, PDB-7cpj: |
化合物 | ![]() ChemComp-ILE: ![]() ChemComp-PRO: |
由来 |
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![]() | ANTIBIOTIC / ribosome |