title: DAZL regulates Tet1 translation in murine embryonic stem cells.
abstract: Embryonic stem cell (ESC) cultures display a heterogeneous gene expression profile, ranging from a pristine naïve pluripotent state to a primed epiblast state. Addition of inhibitors of GSK3β and MEK (so-called 2i conditions) pushes ESC cultures toward a more homogeneous naïve pluripotent state, but the molecular underpinnings of this naïve transition are not completely understood. Here, we demonstrate that DAZL, an RNA-binding protein known to play a key role in germ-cell development, marks a subpopulation of ESCs that is actively transitioning toward naïve pluripotency. Moreover, DAZL plays an essential role in the active reprogramming of cytosine methylation. We demonstrate that DAZL associates with mRNA of Tet1, a catalyst of 5-hydroxylation of methyl-cytosine, and enhances Tet1 mRNA translation. Overexpression of DAZL in heterogeneous ESC cultures results in elevated TET1 protein levels as well as increased global hydroxymethylation. Conversely, null mutation of Dazl severely stunts 2i-mediated TET1 induction and hydroxymethylation. Our results provide insight into the regulation of the acquisition of naïve pluripotency and demonstrate that DAZL enhances TET1-mediated cytosine hydroxymethylation in ESCs that are actively reprogramming to a pluripotent ground state.
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Maaike
authors.initials: M
authors.lastname: Welling
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA Harvard Stem Cell Institute, Cambridge, MA, USA.
authors.forename: Hsu-Hsin
authors.initials: HH
authors.lastname: Chen
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Centre, Utrecht, The Netherlands.
authors.forename: Javier
authors.initials: J
authors.lastname: Muñoz
authors.affiliation: Institute of Molecular Biology, Mainz, Germany.
authors.forename: Michael U
authors.initials: MU
authors.lastname: Musheev
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Lennart
authors.initials: L
authors.lastname: Kester
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Jan Philipp
authors.initials: JP
authors.lastname: Junker
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Centre, Utrecht, The Netherlands.
authors.forename: Nikolai
authors.initials: N
authors.lastname: Mischerikow
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Mandana
authors.initials: M
authors.lastname: Arbab
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Ewart
authors.initials: E
authors.lastname: Kuijk
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA, USA Harvard Stem Cell Institute, Cambridge, MA, USA.
authors.forename: Lev
authors.initials: L
authors.lastname: Silberstein
authors.affiliation: Center for Biomedical Informatics, Harvard Medical School, Boston, MA, USA.
authors.forename: Peter V
authors.initials: PV
authors.lastname: Kharchenko
authors.affiliation: Research Group Reproduction and Genetics, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Mieke
authors.initials: M
authors.lastname: Geens
authors.affiliation: Institute of Molecular Biology, Mainz, Germany Division of Molecular Embryology, DKFZ-ZMBH Alliance, Heidelberg, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Niehrs
authors.affiliation: Research Group Reproduction and Genetics, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Hilde
authors.initials: H
authors.lastname: van de Velde
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Alexander
authors.initials: A
authors.lastname: van Oudenaarden
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands Netherlands Proteomics Centre, Utrecht, The Netherlands.
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, The Netherlands Department of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands n.geijsen@hubrecht.eu.
authors.forename: Niels
authors.initials: N
authors.lastname: Geijsen
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D003596:Cytosine / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D005855:Germ Layers / Q000502:physiology; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D014176:Protein Biosynthesis; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D059467:Transcriptome
chemical_list: C508515:DAZL protein, mouse; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C540839:TET1 protein, mouse; D003596:Cytosine
keywords: 2i conditions; DNA hydroxymethylation; Dazl; TET1; naïve pluripotency