title: Histone variant H3.3 maintains a decondensed chromatin state essential for mouse preimplantation development.
abstract: Histone variants can replace canonical histones in the nucleosome and modify chromatin structure and gene expression. The histone variant H3.3 preferentially associates with active chromatin and has been implicated in the regulation of a diverse range of developmental processes. However, the mechanisms by which H3.3 may regulate gene activity are unclear and gene duplication has hampered an analysis of H3.3 function in mouse. Here, we report that the specific knockdown of H3.3 in fertilized mouse zygotes leads to developmental arrest at the morula stage. This phenotype can be rescued by exogenous H3.3 but not by canonical H3.1 mRNA. Loss of H3.3 leads to over-condensation and mis-segregation of chromosomes as early as the two-cell stage, with corresponding high levels of aneuploidy, but does not appear to affect zygotic gene activation at the two-cell stage or lineage gene transcription at the morula stage. H3.3-deficient embryos have significantly reduced levels of markers of open chromatin, such as H3K36me2 and H4K16Ac. Importantly, a mutation in H3.3K36 that disrupts H3K36 methylation (H3.3K36R) does not rescue the H3.3 knockdown (KD) phenotype. In addition, H3.3 KD embryos have increased incorporation of linker H1. Knockdown of Mof (Kat8), an acetyltransferase specific for H4K16, similarly leads to excessive H1 incorporation. Remarkably, pan-H1 RNA interference (RNAi) partially rescues the chromosome condensation of H3.3 KD embryos and allows development to the blastocyst stage. These results reveal that H3.3 mediates a balance between open and condensed chromatin that is crucial for the fidelity of chromosome segregation during early mouse development.
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Obstetrics and Gynecology and Center for Reproductive Sciences, University of California San Francisco, 35 Medical Center Way, San Francisco, CA 94143, USA.
authors.forename: Chih-Jen
authors.initials: CJ
authors.lastname: Lin
authors.forename: Marco
authors.initials: M
authors.lastname: Conti
authors.forename: Miguel
authors.initials: M
authors.lastname: Ramalho-Santos
mesh_terms: D000818:Animals; D002843:Chromatin / Q000378:metabolism*; D020090:Chromosome Segregation / Q000235:genetics / Q000502:physiology*; D020732:Cytogenetic Analysis; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D055785:Gene Knockdown Techniques; D051548:Histone Acetyltransferases / Q000235:genetics; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D060172:Morpholinos / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D002843:Chromatin; D006657:Histones; D060172:Morpholinos; D034741:RNA, Small Interfering; D051548:Histone Acetyltransferases; C000622683:Kat8 protein, mouse
keywords: Chromatin condensation; Chromosome segregation; Epigenetics; Histone acetylation; Histone methylation; Histone variant H3.3; Linker histone H1; Mouse preimplantation development
references.citation: J Biol Chem. 2006 Jan 6;281(1):559-68
references.pmid: 16267050
references.citation: Genes Dev. 2006 Jul 1;20(13):1744-54
references.pmid: 16818606
references.citation: EMBO J. 2009 Dec 2;28(23):3635-45
references.pmid: 19834459
references.citation: Nat Genet. 2012 Jan 29;44(3):251-3
references.pmid: 22286216
references.citation: Cell. 2001 Nov 2;107(3):323-37
references.pmid: 11701123
references.citation: J Mol Biol. 2008 Sep 12;381(4):816-25
references.pmid: 18653199
references.citation: Cytogenet Genome Res. 2004;104(1-4):131-6
references.pmid: 15162026
references.citation: Nature. 1998 Jul 23;394(6691):369-74
references.pmid: 9690471
references.citation: J Cell Sci. 1994 Mar;107 ( Pt 3):693-701
references.pmid: 8006082
references.citation: Nature. 2005 Oct 27;437(7063):1386-90
references.pmid: 16251970
references.citation: Chromosoma. 2002 Sep;111(3):165-9
references.pmid: 12355205
references.citation: Cell. 2005 Dec 29;123(7):1199-212
references.pmid: 16377562
references.citation: Nature. 2011 Sep 04;477(7366):606-10
references.pmid: 21892189
references.citation: Genes Dev. 2011 Apr 1;25(7):755-66
references.pmid: 21460039
references.citation: Biochim Biophys Acta. 2012 Dec;1822(12):1921-30
references.pmid: 22771499
references.citation: Cell Stem Cell. 2012 Aug 3;11(2):163-78
references.pmid: 22862943
references.citation: Nature. 2012 Jul 5;487(7405):57-63
references.pmid: 22722858
references.citation: Dev Biol. 2005 Jul 1;283(1):40-57
references.pmid: 15975430
references.citation: Int J Dev Biol. 2006;50(5):455-61
references.pmid: 16586346
references.citation: Mol Cell Biol. 2008 Aug;28(16):5093-105
references.pmid: 18541669
references.citation: Nat Struct Mol Biol. 2011 Jun 12;18(7):777-82
references.pmid: 21666677
references.citation: Development. 2003 Sep;130(18):4235-48
references.pmid: 12900441
references.citation: Nature. 2013 Jan 31;493(7434):632-7
references.pmid: 23254936
references.citation: Nat Cell Biol. 2010 Sep;12(9):853-62
references.pmid: 20676102
references.citation: PLoS Genet. 2010 Sep 23;6(9):e1001137
references.pmid: 20885787
references.citation: Cancer Cell. 2012 Oct 16;22(4):425-37
references.pmid: 23079654
references.citation: Nat Genet. 2008 Apr;40(4):411-20
references.pmid: 18311137
references.citation: Cell Rep. 2012 Jun 28;1(6):730-40
references.pmid: 22813747
references.citation: Nature. 2010 Jan 28;463(7280):554-8
references.pmid: 20054296
references.citation: Science. 2011 Apr 22;332(6028):462-5
references.pmid: 21454751
references.citation: Genes Dev. 2010 Jan 15;24(2):159-70
references.pmid: 20040570
references.citation: Cell Res. 2011 Mar;21(3):421-34
references.pmid: 21263457
references.citation: PLoS Genet. 2012 Sep;8(9):e1002938
references.pmid: 23028350
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1525-30
references.pmid: 14732680
references.citation: Nature. 2008 Apr 17;452(7189):877-81
references.pmid: 18354397
references.citation: Biol Reprod. 2010 Aug 1;83(2):177-84
references.pmid: 20410454
references.citation: Curr Biol. 2009 Nov 17;19(21):1816-20
references.pmid: 19781938
references.citation: Mol Cell. 2006 Oct 20;24(2):309-16
references.pmid: 17052464
references.citation: Nature. 2012 Jan 29;482(7384):226-31
references.pmid: 22286061
references.citation: Dev Biol. 2005 Feb 15;278(2):367-80
references.pmid: 15680357
references.citation: Cell. 2010 Mar 5;140(5):678-91
references.pmid: 20211137
references.citation: Mol Cell. 2002 Jun;9(6):1191-200
references.pmid: 12086617
references.citation: Science. 2006 Feb 10;311(5762):844-7
references.pmid: 16469925
references.citation: Mol Cell. 2004 Oct 8;16(1):93-105
references.pmid: 15469825
references.citation: Proc Natl Acad Sci U S A. 2011 Jan 4;108(1):85-90
references.pmid: 21173268
references.citation: Proc Natl Acad Sci U S A. 2006 Apr 25;103(17):6428-35
references.pmid: 16571659
references.citation: Mol Cell Biol. 2008 Jan;28(1):397-409
references.pmid: 17967868
references.citation: EMBO J. 2007 Dec 12;26(24):4974-84
references.pmid: 18007591
references.citation: Development. 2013 Feb;140(4):853-60
references.pmid: 23318639