title: Endogenous Mouse Dicer Is an Exclusively Cytoplasmic Protein.
abstract: Dicer is a large multi-domain protein responsible for the ultimate step of microRNA and short-interfering RNA biogenesis. In human and mouse cell lines, Dicer has been shown to be important in the nuclear clearance of dsRNA as well as the establishment of chromatin modifications. Here we set out to unambiguously define the cellular localization of Dicer in mice to understand if this is a conserved feature of mammalian Dicer in vivo. To this end, we utilized an endogenously epitope tagged Dicer knock-in mouse allele. From primary mouse cell lines and adult tissues, we determined with certainty by biochemical fractionation and confocal immunofluorescence microscopy that endogenous Dicer is exclusively cytoplasmic. We ruled out the possibility that a fraction of Dicer shuttles to and from the nucleus as well as that FGF or DNA damage signaling induce Dicer nuclear translocation. We also explored Dicer localization during the dynamic and developmental context of embryogenesis, where Dicer is ubiquitously expressed and strictly cytoplasmic in all three germ layers as well as extraembryonic tissues. Our data exclude a direct role for Dicer in the nuclear RNA processing in the mouse.
authors.affiliation: Mouse Biology Unit, European Molecular Biology Laboratory (EMBL), Monterotondo, Italy.|MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
authors.forename: Christian
authors.initials: C
authors.lastname: Much
authors.affiliation: Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
authors.forename: Tania
authors.identifier: 0000-0002-7748-2939
authors.initials: T
authors.lastname: Auchynnikava
authors.affiliation: Genomics Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors.forename: Dinko
authors.initials: D
authors.lastname: Pavlinic
authors.affiliation: Mouse Biology Unit, European Molecular Biology Laboratory (EMBL), Monterotondo, Italy.
authors.forename: Andreas
authors.initials: A
authors.lastname: Buness
authors.affiliation: Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
authors.forename: Juri
authors.initials: J
authors.lastname: Rappsilber
authors.affiliation: Genomics Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Benes
authors.affiliation: Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
authors.forename: Robin
authors.initials: R
authors.lastname: Allshire
authors.affiliation: Mouse Biology Unit, European Molecular Biology Laboratory (EMBL), Monterotondo, Italy.|MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
authors.forename: Dónal
authors.initials: D
authors.lastname: O'Carroll
mesh_terms: D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000235:genetics; D012330:RNA, Double-Stranded / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism*
chemical_list: D035683:MicroRNAs; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases