[en] Nonsense-Mediated mRNA Decay (NMD) is a key control mechanism of RNA quality widely described to target mRNA harbouring Premature Termination Codon (PTC). However, recent studies suggested the existence of non-canonical pathways which remain unresolved. One of these alternative pathways suggested that specific mRNA could be targeted through their 3' UTR (Untranslated Region), which contain various elements involved in mRNA stability regulation. This study focused on 3'UTR of mRNA encoding NMD factors, on which we observed an enrichment of binding sites for UPF1 and eIF4A3 proteins, two important NMD factors. Using GFP reporter constructs containing the 3'UTR of these NMD mRNA fused to the GFP cDNA, we showed that GFP expression was significantly increased upon eIF4A3 inhibition, suggesting mRNA level stabilization. Furthermore, co-immunoprecipitation targeting UPF1 revealed that its interaction with mRNA encoding NMD factors was disrupted when cells were previously treated with the eIF4A3 inhibitor. We therefore propose that eIF4A3 might be necessary to recruit UPF1 and trigger the degradation of these transcripts through a non-canonical 3'UTR-dependent mechanism.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Mercier, Chloé; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France
Durand, Jules ; Université de Liège - ULiège > Département de pharmacie ; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France
Fraichard, Annick; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France
Perez, Valérie; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France
Hervouet, Eric; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France, EPIGENExp platform, Université Franche-Comté, F-25000, Besançon, France
Peixoto, Paul ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > GIGA-R : Labo de recherche sur les métastases ; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France, EPIGENExp platform, Université Franche-Comté, F-25000, Besançon, France
Delage-Mourroux, Regis; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France
Guittaut, Michaël ; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France
Baguet, Aurélie ; Université Franche-Comté, INSERM, EFS BFC, UMR1098, Interactions Hôte-Greffon-Tumeur/Ingénierie Cellulaire et Génique, F-25000, Besançon, France. Electronic address: aurelie.baguet@univ-fcomte.fr
Language :
English
Title :
Chemical inhibition of eIF4A3 abolishes UPF1 recruitment onto mRNA encoding NMD factors and restores their expression.
Publication date :
02 February 2025
Journal title :
Biochemical and Biophysical Research Communications
Chlo\u00E9 Mercier and Jules Durand were supported by a fellowship from the MESR \u201CMinist\u00E8re de l\u2019Enseignement Sup\u00E9rieur et de la Recherche.\u201D This work was supported by fundings from Ligue Contre le Cancer (CCIR Est 2020, CCIR Est 2021 and CCIR Est 2024), INSERM, EFS BFC, Univ. Bourgogne Franche-Comt\u00E9 and the \u201CR\u00E9gion Bourgogne Franche-Comt\u00E9\u201D (Convention Amor\u00E7age 2021Y-08309). We thank the DIMAcell platform for their technical support during Incucyte experiments. We thank Dr. M\u00FChlemann and collaborators for sending us supplementary data from their previous publicationChlo\u00E9 Mercier and Jules Durand were supported by a fellowship from the MESR \u201CMinist\u00E8re de l'Enseignement Sup\u00E9rieur et de la Recherche.\u201D This work was supported by fundings from Ligue Contre le Cancer (CCIR Est 2020, CCIR Est 2021 and CCIR Est 2024), INSERM, EFS BFC, Univ. Bourgogne Franche-Comt\u00E9 and the \u201CR\u00E9gion Bourgogne Franche-Comt\u00E9\u201D (Convention Amor\u00E7age 2021Y-08309). We thank the DIMAcell platform for their technical support during Incucyte experiments. We thank Dr. M\u00FChlemann and collaborators for sending us supplementary data from their previous publication.
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