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000296113 1001_ $$00000-0003-3115-2501$$aMeurs, Romane$$b0
000296113 245__ $$aMCTS2 and distinct eIF2D roles in uORF-dependent translation regulation revealed by in vitro re-initiation assays.
000296113 260__ $$aHoboken, NJ [u.a.]$$bWiley$$c2025
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000296113 520__ $$aRibosomes 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.
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000296113 650_7 $$2Other$$aDENR-MCTS1
000296113 650_7 $$2Other$$aIn Vitro Translation
000296113 650_7 $$2Other$$aRe-Initiation
000296113 650_7 $$2Other$$aeIF2D
000296113 650_7 $$2Other$$auORF
000296113 7001_ $$aDe Matos, Mara$$b1
000296113 7001_ $$00009-0007-2710-1273$$aBothe, Adrian$$b2
000296113 7001_ $$00000-0001-6023-0519$$aGuex, Nicolas$$b3
000296113 7001_ $$0P:(DE-He78)92828afe777c83bb1efb2f2f5bbb060a$$aWeber, Tobias$$b4$$udkfz
000296113 7001_ $$0P:(DE-He78)5ebc16fd8019dbfde58e0125b001b599$$aTeleman, Aurelio$$b5$$udkfz
000296113 7001_ $$00000-0002-9527-210X$$aBan, Nenad$$b6
000296113 7001_ $$00000-0001-5114-2824$$aGatfield, David$$b7
000296113 773__ $$0PERI:(DE-600)1467419-1$$a10.1038/s44318-024-00347-3$$n3$$p854-876$$tThe EMBO journal$$v44$$x0261-4189$$y2025
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