Datasets

Title	Assembly and functionality of the ribosome with tethered subunits
Organism
Number of Samples	6
Release Date	2018/09/04 00:00
Sequencing Types
Protocol Details

Title
Authors	Aleksashin NA,Leppik M,Hockenberry AJ,Klepacki D,Vázquez-Laslop N,Jewett MC,Remme J,Mankin AS
Journal	Nature communications
Publication Date	2019 Feb 25
Abstract	Ribo-T is an engineered ribosome whose small and large subunits are tethered together by linking 16S rRNA and 23S rRNA in a single molecule. Although Ribo-T can support cell proliferation in the absence of wild type ribosomes, Ribo-T cells grow slower than those with wild type ribosomes. Here, we show that cell growth defect is likely explained primarily by slow Ribo-T assembly rather than its imperfect functionality. Ribo-T maturation is stalled at a late assembly stage. Several post-transcriptional rRNA modifications and some ribosomal proteins are underrepresented in the accumulated assembly intermediates and rRNA ends are incompletely trimmed. Ribosome profiling of Ribo-T cells shows no defects in translation elongation but reveals somewhat higher occupancy by Ribo-T of the start codons and to a lesser extent stop codons, suggesting that subunit tethering mildly affects the initiation and termination stages of translation. Understanding limitations of Ribo-T system offers ways for its future development.
PMC	PMC6389949
PMID	30804338
DOI

Aleksashin et al. 2019 (PRJNA489275)