title: Tet1 and Tet2 regulate 5-hydroxymethylcytosine production and cell lineage specification in mouse embryonic stem cells.
abstract: TET family enzymes convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) in DNA. Here, we show that Tet1 and Tet2 are Oct4-regulated enzymes that together sustain 5hmC in mouse embryonic stem cells (ESCs) and are induced concomitantly with 5hmC during reprogramming of fibroblasts to induced pluripotent stem cells. ESCs depleted of Tet1 by RNAi show diminished expression of the Nodal antagonist Lefty1 and display hyperactive Nodal signaling and skewed differentiation into the endoderm-mesoderm lineage in embryoid bodies in vitro. In Fgf4- and heparin-supplemented culture conditions, Tet1-depleted ESCs activate the trophoblast stem cell lineage determinant Elf5 and can colonize the placenta in midgestation embryo chimeras. Consistent with these findings, Tet1-depleted ESCs form aggressive hemorrhagic teratomas with increased endoderm, reduced neuroectoderm, and ectopic appearance of trophoblastic giant cells. Thus, 5hmC is an epigenetic modification associated with the pluripotent state, and Tet1 functions to regulate the lineage differentiation potential of ESCs.
authors.affiliation: Immune Disease Institute and Program in Cellular and Molecular Medicine, Children's Hospital Boston, Boston, MA 02115, USA.
authors.forename: Kian Peng
authors.initials: KP
authors.lastname: Koh
authors.forename: Akiko
authors.initials: A
authors.lastname: Yabuuchi
authors.forename: Sridhar
authors.initials: S
authors.lastname: Rao
authors.forename: Yun
authors.initials: Y
authors.lastname: Huang
authors.forename: Kerrianne
authors.initials: K
authors.lastname: Cunniff
authors.forename: Julie
authors.initials: J
authors.lastname: Nardone
authors.forename: Asta
authors.initials: A
authors.lastname: Laiho
authors.forename: Mamta
authors.initials: M
authors.lastname: Tahiliani
authors.forename: Cesar A
authors.initials: CA
authors.lastname: Sommer
authors.forename: Gustavo
authors.initials: G
authors.lastname: Mostoslavsky
authors.forename: Riitta
authors.initials: R
authors.lastname: Lahesmaa
authors.forename: Stuart H
authors.initials: SH
authors.lastname: Orkin
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Rodig
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
authors.forename: Anjana
authors.initials: A
authors.lastname: Rao
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000818:Animals; D001665:Binding Sites / Q000235:genetics / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D019070:Cell Lineage; D047369:Chromatin Immunoprecipitation; D019295:Computational Biology; D003596:Cytosine / Q000031:analogs & derivatives* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D013724:Teratoma / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011518:Proto-Oncogene Proteins; C540839:TET1 protein, mouse; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D049308:Dioxygenases; C551914:Tet2 protein, mouse