title: Suv4-20h histone methyltransferases promote neuroectodermal differentiation by silencing the pluripotency-associated Oct-25 gene.
abstract: Post-translational modifications (PTMs) of histones exert fundamental roles in regulating gene expression. During development, groups of PTMs are constrained by unknown mechanisms into combinatorial patterns, which facilitate transitions from uncommitted embryonic cells into differentiated somatic cell lineages. Repressive histone modifications such as H3K9me3 or H3K27me3 have been investigated in detail, but the role of H4K20me3 in development is currently unknown. Here we show that Xenopus laevis Suv4-20h1 and h2 histone methyltransferases (HMTases) are essential for induction and differentiation of the neuroectoderm. Morpholino-mediated knockdown of the two HMTases leads to a selective and specific downregulation of genes controlling neural induction, thereby effectively blocking differentiation of the neuroectoderm. Global transcriptome analysis supports the notion that these effects arise from the transcriptional deregulation of specific genes rather than widespread, pleiotropic effects. Interestingly, morphant embryos fail to repress the Oct4-related Xenopus gene Oct-25. We validate Oct-25 as a direct target of xSu4-20h enzyme mediated gene repression, showing by chromatin immunoprecipitaton that it is decorated with the H4K20me3 mark downstream of the promoter in normal, but not in double-morphant, embryos. Since knockdown of Oct-25 protein significantly rescues the neural differentiation defect in xSuv4-20h double-morphant embryos, we conclude that the epistatic relationship between Suv4-20h enzymes and Oct-25 controls the transit from pluripotent to differentiation-competent neural cells. Consistent with these results in Xenopus, murine Suv4-20h1/h2 double-knockout embryonic stem (DKO ES) cells exhibit increased Oct4 protein levels before and during EB formation, and reveal a compromised and biased capacity for in vitro differentiation, when compared to normal ES cells. Together, these results suggest a regulatory mechanism, conserved between amphibians and mammals, in which H4K20me3-dependent restriction of specific POU-V genes directs cell fate decisions, when embryonic cells exit the pluripotent state.
authors: Adolf Butenandt Institut, Institut für Molekularbiologie, Ludwig Maximilians-Universität, München, Germany.
authors: Dario
authors: D
authors: Nicetto
authors: Matthias
authors: M
authors: Hahn
authors: Julia
authors: J
authors: Jung
authors: Tobias D
authors: TD
authors: Schneider
authors: Tobias
authors: T
authors: Straub
authors: Robert
authors: R
authors: David
authors: Gunnar
authors: G
authors: Schotta
authors: Ralph A W
authors: RA
authors: Rupp
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D002843:Chromatin / Q000235:genetics; D019175:DNA Methylation; D004625:Embryo, Nonmammalian; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics* / Q000378:metabolism; D054258:Neural Plate / Q000254:growth & development / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D050809:POU Domain Factors / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D029867:Xenopus Proteins / Q000235:genetics* / Q000378:metabolism; D014982:Xenopus laevis / Q000235:genetics / Q000254:growth & development
chemical_list: D002843:Chromatin; D050814:Octamer Transcription Factor-3; D050809:POU Domain Factors; C494174:Pou5f1 protein, mouse; C570043:Suv4-20h protein, mouse; D029867:Xenopus Proteins; C527329:pou5f3.2 protein, Xenopus; D011495:Histone-Lysine N-Methyltransferase; C581298:KMT5B protein, Xenopus; C581299:KMT5C protein, Xenopus
references: J Exp Med. 2010 Oct 25;207(11):2287-95
references: 20975044
references: Genesis. 2004 Jan;38(1):32-8
references: 14755802
references: Mech Dev. 1996 Mar;55(1):19-32
references: 8734496
references: Development. 2007 Feb;134(4):635-46
references: 17215298
references: Heredity (Edinb). 2010 Jul;105(1):24-37
references: 20442736
references: Genes Dev. 2004 Jun 1;18(11):1251-62
references: 15145825
references: Proc Natl Acad Sci U S A. 2011 Feb 22;108(8):3116-23
references: 21282610
references: Genes Dev. 2002 Jul 15;16(14):1779-91
references: 12130538
references: Dev Cell. 2009 Sep;17(3):425-34
references: 19758566
references: J Biol Chem. 2008 Dec 5;283(49):34168-77
references: 18922797
references: Cell. 2010 Jun 11;141(6):943-55
references: 20550931
references: Dev Biol. 2004 Dec 15;276(2):337-51
references: 15581869
references: Cell. 2005 Sep 23;122(6):947-56
references: 16153702
references: Oncogene. 2002 Jan 3;21(1):148-52
references: 11791185
references: Mech Dev. 2007 Nov-Dec;124(11-12):840-55
references: 17950579
references: Genome Biol. 2005;6(8):227
references: 16086857
references: Nat Rev Mol Cell Biol. 2009 Aug;10(8):526-37
references: 19603040
references: Nature. 2007 Aug 2;448(7153):553-60
references: 17603471
references: Cell. 2011 May 13;145(4):555-70
references: 21529909
references: Nat Genet. 2003 Jun;34(2):187-92
references: 12740577
references: PLoS One. 2011;6(7):e22548
references: 21814581
references: Nature. 2010 Apr 8;464(7290):922-6
references: 20336069
references: Nat Struct Mol Biol. 2007 Nov;14(11):1008-16
references: 17984963
references: Dev Cell. 2010 Nov 16;19(5):662-74
references: 21074717
references: Mol Cell Biol. 1992 Feb;12(2):638-49
references: 1732736
references: Curr Opin Genet Dev. 2004 Apr;14(2):155-64
references: 15196462
references: Birth Defects Res C Embryo Today. 2009 Sep;87(3):249-62
references: 19750523
references: Stem Cell Rev Rep. 2010 Mar;6(1):62-73
references: 20148317
references: Genes Dev. 2011 Feb 1;25(3):238-50
references: 21245162
references: PLoS Genet. 2012;8(5):e1002691
references: 22589736
references: Cell. 2007 May 18;129(4):823-37
references: 17512414
references: Mech Dev. 2006 Aug;123(8):614-25
references: 16860542
references: EMBO J. 2003 Jan 15;22(2):292-303
references: 12514135
references: Nat Rev Mol Cell Biol. 2006 Jul;7(7):540-6
references: 16723974
references: Nature. 2000 Aug 10;406(6796):593-9
references: 10949293
references: Nat Cell Biol. 2008 Mar;10(3):338-45
references: 18297060
references: J Cell Biochem. 2009 Sep 1;108(1):22-34
references: 19623574
references: Genes Dev. 2008 Aug 1;22(15):2048-61
references: 18676810
references: Dev Dyn. 2009 Jun;238(6):1422-32
references: 19334278
references: Dev Cell. 2006 Jan;10(1):105-16
references: 16399082
references: J Biol Chem. 2004 Oct 15;279(42):43735-43
references: 15292233
references: Mol Cell Biol. 2011 Apr;31(8):1594-609
references: 21321083
references: Nature. 2006 May 18;441(7091):349-53
references: 16625203
references: Development. 2006 May;133(10):2011-22
references: 16651543
references: Nat Rev Neurosci. 2002 Apr;3(4):271-80
references: 11967557
references: EMBO J. 2006 Aug 9;25(15):3664-74
references: 16858397
references: Science. 2010 Apr 30;328(5978):633-6
references: 20431018
references: Neuron. 1991 Apr;6(4):499-515
references: 1901716
references: Cell. 2011 Jun 10;145(6):875-89
references: 21663792