title: Differential utilization of decapping enzymes in mammalian mRNA decay pathways.
abstract: mRNA decapping is a crucial step in the regulation of mRNA stability and gene expression. Dcp2 is an mRNA decapping enzyme that has been widely studied. We recently reported the presence of a second mammalian cytoplasmic decapping enzyme, Nudt16. Here we address the differential utilization of the two decapping enzymes in specified mRNA decay processes. Using mouse embryonic fibroblast (MEF) cell lines derived from a hypomorphic knockout of the Dcp2 gene with undetectable levels of Dcp2 or MEF cell lines harboring a Nudt16-directed shRNA to generate reduced levels of Nudt16, we demonstrate the distinct roles for Dcp2 and Nudt16 in nonsense-mediated mRNA decay (NMD), decay of ARE-containing mRNA and miRNA-mediated silencing. Our results indicated that NMD preferentially utilizes Dcp2 rather than Nudt16; Dcp2 and Nudt16 are redundant in miRNA-mediated silencing; and Dcp2 and Nudt16 are differentially utilized for ARE-mRNA decay. These data demonstrate that the two distinct decapping enzymes can uniquely function in specific mRNA decay processes in mammalian cells.
authors.affiliation: Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, New Jersey 08854-8082, USA.
authors.forename: You
authors.initials: Y
authors.lastname: Li
authors.forename: Mangen
authors.initials: M
authors.lastname: Song
authors.forename: Megerditch
authors.initials: M
authors.lastname: Kiledjian
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D004722:Endoribonucleases / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors / Q000502:physiology*; D012315:RNA Caps / Q000235:genetics*; D020871:RNA Stability / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D035683:MicroRNAs; D012315:RNA Caps; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C030052:mRNA decapping enzymes; D008156:Luciferases; C554183:Dcp2 protein, mouse; D004722:Endoribonucleases; C549897:NUDT16 protein, mouse; D011755:Pyrophosphatases