%0 Journal Article
%A Meurs, Romane
%A De Matos, Mara
%A Bothe, Adrian
%A Guex, Nicolas
%A Weber, Tobias
%A Teleman, Aurelio
%A Ban, Nenad
%A Gatfield, David
%T MCTS2 and distinct eIF2D roles in uORF-dependent translation regulation revealed by in vitro re-initiation assays.
%J The EMBO journal
%V 44
%N 3
%@ 0261-4189
%C Hoboken, NJ [u.a.]
%I Wiley
%M DKFZ-2025-00043
%P 854-876
%D 2025
%Z 2025 Feb;44(3):854-876
%X Ribosomes scanning from the mRNA 5' cap to the start codon may initiate at upstream open reading frames (uORFs), decreasing protein biosynthesis. Termination at a uORF can lead to re-initiation, where 40S subunits resume scanning and initiate another translation event downstream. The noncanonical translation factors MCTS1-DENR participate in re-initiation at specific uORFs, but knowledge of other trans-acting factors or uORF features influencing re-initiation is limited. Here, we establish a cell-free re-initiation assay using HeLa lysates to address this question. Comparing in vivo and in vitro re-initiation on uORF-containing reporters, we validate MCTS1-DENR-dependent re-initiation in vitro. Using this system and ribosome profiling in cells, we found that knockdown of the MCTS1-DENR homolog eIF2D causes widespread gene deregulation unrelated to uORF translation, and thus distinct to MCTS1-DENR-dependent re-initiation regulation. Additionally, we identified MCTS2, encoded by an Mcts1 retrogene, as a DENR partner promoting re-initiation in vitro, providing a plausible explanation for clinical differences associated with DENR vs. MCTS1 mutations in humans.
%K DENR-MCTS1 (Other)
%K In Vitro Translation (Other)
%K Re-Initiation (Other)
%K eIF2D (Other)
%K uORF (Other)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:39748120
%R 10.1038/s44318-024-00347-3
%U https://inrepo02.dkfz.de/record/296113