Datasets

Title	Selective Translation Complex Profiling in yeast and human Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes [Yeast]
Organism
Number of Samples	15
Release Date	2019/10/20 00:00
Sequencing Types
Protocol Details

Title
Authors	Wagner S,Herrmannová A,Hronová V,Gunišová S,Sen ND,Hannan RD,Hinnebusch AG,Shirokikh NE,Preiss T,Valášek LS
Journal	Molecular cell
Publication Date	2020 Aug 20
Abstract	Translational control targeting the initiation phase is central to the regulation of gene expression. Understanding all of its aspects requires substantial technological advancements. Here we modified yeast translation complex profile sequencing (TCP-seq), related to ribosome profiling, and adapted it for mammalian cells. Human TCP-seq, capable of capturing footprints of 40S subunits (40Ss) in addition to 80S ribosomes (80Ss), revealed that mammalian and yeast 40Ss distribute similarly across 5'TRs, indicating considerable evolutionary conservation. We further developed yeast and human selective TCP-seq (Sel-TCP-seq), enabling selection of 40Ss and 80Ss associated with immuno-targeted factors. Sel-TCP-seq demonstrated that eIF2 and eIF3 travel along 5' UTRs with scanning 40Ss to successively dissociate upon AUG recognition; notably, a proportion of eIF3 lingers on during the initial elongation cycles. Highlighting Sel-TCP-seq versatility, we also identified four initiating 48S conformational intermediates, provided novel insights into ATF4 and GCN4 mRNA translational control, and demonstrated co-translational assembly of initiation factor complexes. Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.
PMC	PMC7447980
PMID	32589964
DOI

Wagner et al. 2020 (PRJNA578566)