Gastrulation initiates with the formation of the primitive streak, during which, cells of the epiblast delaminate to form the mesoderm and definitive endoderm. At this stage, the pluripotent cell population of the epiblast undergoes very rapid proliferation and extensive epigenetic programming. Here we show that Fam208a, a new epigenetic modifier, is essential for early post-implantation development. We show that Fam208a mutation leads to impaired primitive streak elongation and delayed epithelial-to-mesenchymal transition. Fam208a mutant epiblasts had increased expression of p53 pathway genes as well as several pluripotency-associated long non-coding RNAs. Fam208a mutants exhibited an increase in p53-driven apoptosis and complete removal of p53 could partially rescue their gastrulation block. This data demonstrates a new in vivo function of Fam208a in maintaining epiblast fitness, establishing it as an important factor at the onset of gastrulation when cells are exiting pluripotency.

Czech Centre for Phenogenomics Division BIOCEV Institute of Molecular Genetics of the CAS v v i Vestec Czech Republic

Department of Physiology Faculty of Medicine University of Toronto Ontario Canada

Faculty of Science Charles University Prague Czech Republic

Laboratory of Cell and Developmental Biology Institute of Molecular Genetics of the CAS v v i Krc Czech Republic

Laboratory of Genomics and Bioinformatics Institute of Molecular Genetics of the CAS v v i Krc Czech Republic

Laboratory of Transgenic Models of Diseases Division BIOCEV Institute of Molecular Genetics of the CAS v v i Vestec Czech Republic

Erratum In

Sci Rep. 2018 Apr 5;8(1):5762. doi: 10.1038/s41598-018-22650-w. PubMed

See more in PubMed

Snow MHL. Gastrulation in the mouse: Growth and regionalization of the epiblast. J. Embryol. exp. Morph. 1977;42:293–303.

Lewis NE, Rossant J. Mechanism of size regulation in mouse embryo aggregates. J Embryol Exp Morphol. 1982;72:169–181. PubMed

Power MA, Tam PP. Onset of gastrulation, morphogenesis and somitogenesis in mouse embryos displaying compensatory growth. Anat Embryol (Berl) 1993;187:493–504. doi: 10.1007/BF00174425. PubMed DOI

Arnold SJ, Robertson EJ. Making a commitment: cell lineage allocation and axis patterning in the early mouse embryo. Nat Rev Mol Cell Biol. 2009;10:91–103. doi: 10.1038/nrm2618. PubMed DOI

Tam PP, Loebel DA. Gene function in mouse embryogenesis: get set for gastrulation. Nat Rev Genet. 2007;8:368–381. doi: 10.1038/nrg2084. PubMed DOI

Blewitt ME, et al. An N-ethyl-N-nitrosourea screen for genes involved in variegation in the mouse. Proc Natl Acad Sci USA. 2005;102:7629–7634. doi: 10.1073/pnas.0409375102. PubMed DOI PMC

Daxinger L, et al. An ENU mutagenesis screen identifies novel and known genes involved in epigenetic processes in the mouse. Genome Biol. 2013;14:R96. doi: 10.1186/gb-2013-14-9-r96. PubMed DOI PMC

Whitelaw NC, et al. Reduced levels of two modifiers of epigenetic gene silencing, Dnmt3a and Trim28, cause increased phenotypic noise. Genome Biol. 2010;11:R111. doi: 10.1186/gb-2010-11-11-r111. PubMed DOI PMC

Isbel L, et al. Trim33 Binds and Silences a Class of Young Endogenous Retroviruses in the Mouse Testis; a Novel Component of the Arms Race between Retrotransposons and the Host Genome. PLoS Genet. 2015;11:e1005693. doi: 10.1371/journal.pgen.1005693. PubMed DOI PMC

Harten SK, et al. The first mouse mutants of D14Abb1e (Fam208a) show that it is critical for early development. Mamm Genome. 2014;25:293–303. doi: 10.1007/s00335-014-9516-0. PubMed DOI PMC

Tchasovnikarova IA, et al. GENE SILENCING. Epigenetic silencing by the HUSH complex mediates position-effect variegation in human cells. Science. 2015;348:1481–1485. doi: 10.1126/science.aaa7227. PubMed DOI PMC

Timms RT, Tchasovnikarova IA, Lehner PJ. Position-effect variegation revisited: HUSHing up heterochromatin in human cells. BioEssays: news and reviews in molecular, cellular and developmental biology. 2016;38:333–343. doi: 10.1002/bies.201500184. PubMed DOI

Lachner M, Jenuwein T. The many faces of histone lysine methylation. Curr Opin Cell Biol. 2002;14:286–298. doi: 10.1016/S0955-0674(02)00335-6. PubMed DOI

Young RA. Control of the embryonic stem cell state. Cell. 2011;144:940–954. doi: 10.1016/j.cell.2011.01.032. PubMed DOI PMC

Xi Q, et al. A poised chromatin platform for TGF-beta access to master regulators. Cell. 2011;147:1511–1524. doi: 10.1016/j.cell.2011.11.032. PubMed DOI PMC

Dodge JE, Kang YK, Beppu H, Lei H, Li E. Histone H3-K9 methyltransferase ESET is essential for early development. Mol Cell Biol. 2004;24:2478–2486. doi: 10.1128/MCB.24.6.2478-2486.2004. PubMed DOI PMC

Morsut L, et al. Negative control of Smad activity by ectodermin/Tif1gamma patterns the mammalian embryo. Development. 2010;137:2571–2578. doi: 10.1242/dev.053801. PubMed DOI

Cammas F, et al. Mice lacking the transcriptional corepressor TIF1beta are defective in early postimplantation development. Development. 2000;127:2955–2963. PubMed

Beck S, et al. Extraembryonic proteases regulate Nodal signalling during gastrulation. Nat Cell Biol. 2002;4:981–985. doi: 10.1038/ncb890. PubMed DOI

Donnison M, et al. Loss of the extraembryonic ectoderm in Elf5 mutants leads to defects in embryonic patterning. Development. 2005;132:2299–2308. doi: 10.1242/dev.01819. PubMed DOI

Fujiwara T, Dunn NR, Hogan BL. Bone morphogenetic protein 4 in the extraembryonic mesoderm is required for allantois development and the localization and survival of primordial germ cells in the mouse. Proc Natl Acad Sci USA. 2001;98:13739–13744. doi: 10.1073/pnas.241508898. PubMed DOI PMC

Winnier G, Blessing M, Labosky PA, Hogan BL. Bone morphogenetic protein-4 is required for mesoderm formation and patterning in the mouse. Genes Dev. 1995;9:2105–2116. doi: 10.1101/gad.9.17.2105. PubMed DOI

Herrmann BG. Expression pattern of the Brachyury gene in whole-mount TWis/TWis mutant embryos. Development. 1991;113:913–917. PubMed

Ding J, et al. Cripto is required for correct orientation of the anterior-posterior axis in the mouse embryo. Nature. 1998;395:702–707. doi: 10.1038/27215. PubMed DOI

Abdelkhalek HB, et al. The mouse homeobox gene Not is required for caudal notochord development and affected by the truncate mutation. Genes Dev. 2004;18:1725–1736. doi: 10.1101/gad.303504. PubMed DOI PMC

Yamanaka Y, Tamplin OJ, Beckers A, Gossler A, Rossant J. Live imaging and genetic analysis of mouse notochord formation reveals regional morphogenetic mechanisms. Dev Cell. 2007;13:884–896. doi: 10.1016/j.devcel.2007.10.016. PubMed DOI

Echelard Y, et al. Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity. Cell. 1993;75:1417–1430. doi: 10.1016/0092-8674(93)90627-3. PubMed DOI

Monaghan AP, Kaestner KH, Grau E, Schutz G. Postimplantation expression patterns indicate a role for the mouse forkhead/HNF-3 alpha, beta and gamma genes in determination of the definitive endoderm, chordamesoderm and neuroectoderm. Development. 1993;119:567–578. PubMed

Sasaki H, Hogan BL. Differential expression of multiple fork head related genes during gastrulation and axial pattern formation in the mouse embryo. Development. 1993;118:47–59. PubMed

Ang SL, et al. The formation and maintenance of the definitive endoderm lineage in the mouse: involvement of HNF3/forkhead proteins. Development. 1993;119:1301–1315. PubMed

Burtscher I, Lickert H. Foxa2 regulates polarity and epithelialization in the endoderm germ layer of the mouse embryo. Development. 2009;136:1029–1038. doi: 10.1242/dev.028415. PubMed DOI

Burdsal CA, Damsky CH, Pedersen RA. The role of E-cadherin and integrins in mesoderm differentiation and migration at the mammalian primitive streak. Development. 1993;118:829–844. PubMed

Ciruna B, Rossant J. FGF signaling regulates mesoderm cell fate specification and morphogenetic movement at the primitive streak. Dev Cell. 2001;1:37–49. doi: 10.1016/S1534-5807(01)00017-X. PubMed DOI

Damjanov I, Damjanov A, Damsky CH. Developmentally regulated expression of the cell-cell adhesion glycoprotein cell-CAM 120/80 in peri-implantation mouse embryos and extraembryonic membranes. Developmental biology. 1986;116:194–202. doi: 10.1016/0012-1606(86)90056-4. PubMed DOI

Smith DE, Franco del Amo F, Gridley T. Isolation of Sna, a mouse gene homologous to the Drosophila genes snail and escargot: its expression pattern suggests multiple roles during postimplantation development. Development. 1992;116:1033–1039. PubMed

Yamaguchi TP, Harpal K, Henkemeyer M, Rossant J. fgfr-1 is required for embryonic growth and mesodermal patterning during mouse gastrulation. Genes Dev. 1994;8:3032–3044. doi: 10.1101/gad.8.24.3032. PubMed DOI

Sun X, Meyers EN, Lewandoski M, Martin GR. Targeted disruption of Fgf8 causes failure of cell migration in the gastrulating mouse embryo. Genes Dev. 1999;13:1834–1846. doi: 10.1101/gad.13.14.1834. PubMed DOI PMC

Brennan J, et al. Nodal signalling in the epiblast patterns the early mouse embryo. Nature. 2001;411:965–969. doi: 10.1038/35082103. PubMed DOI

Arnold SJ, Hofmann UK, Bikoff EK, Robertson EJ. Pivotal roles for eomesodermin during axis formation, epithelium-to-mesenchyme transition and endoderm specification in the mouse. Development. 2008;135:501–511. doi: 10.1242/dev.014357. PubMed DOI PMC

Ciruna BG, Rossant J. Expression of the T-box gene Eomesodermin during early mouse development. Mech Dev. 1999;81:199–203. doi: 10.1016/S0925-4773(98)00243-3. PubMed DOI

Hancock SN, Agulnik SI, Silver LM, Papaioannou VE. Mapping and expression analysis of the mouse ortholog of Xenopus Eomesodermin. Mech Dev. 1999;81:205–208. doi: 10.1016/S0925-4773(98)00244-5. PubMed DOI

Russ AP, et al. Eomesodermin is required for mouse trophoblast development and mesoderm formation. Nature. 2000;404:95–99. doi: 10.1038/35003601. PubMed DOI

Belo JA, et al. Cerberus-like is a secreted BMP and nodal antagonist not essential for mouse development. Genesis. 2000;26:265–270. doi: 10.1002/(SICI)1526-968X(200004)26:4<265::AID-GENE80>3.0.CO;2-4. PubMed DOI

Belo JA, et al. Cerberus-like is a secreted factor with neutralizing activity expressed in the anterior primitive endoderm of the mouse gastrula. Mech Dev. 1997;68:45–57. doi: 10.1016/S0925-4773(97)00125-1. PubMed DOI

Biben C, et al. Murine cerberus homologue mCer-1: a candidate anterior patterning molecule. Developmental biology. 1998;194:135–151. doi: 10.1006/dbio.1997.8812. PubMed DOI

Pearce JJ, Penny G, Rossant J. A mouse cerberus/Dan-related gene family. Developmental biology. 1999;209:98–110. doi: 10.1006/dbio.1999.9240. PubMed DOI

Barnes JD, Crosby JL, Jones CM, Wright CV, Hogan BL. Embryonic expression of Lim-1, the mouse homolog of Xenopus Xlim-1, suggests a role in lateral mesoderm differentiation and neurogenesis. Developmental biology. 1994;161:168–178. doi: 10.1006/dbio.1994.1018. PubMed DOI

Tsang TE, et al. Lim1 activity is required for intermediate mesoderm differentiation in the mouse embryo. Developmental biology. 2000;223:77–90. doi: 10.1006/dbio.2000.9733. PubMed DOI

Simeone A, et al. A vertebrate gene related to orthodenticle contains a homeodomain of the bicoid class and demarcates anterior neuroectoderm in the gastrulating mouse embryo. EMBO J. 1993;12:2735–2747. PubMed PMC

Ang SL, Conlon RA, Jin O, Rossant J. Positive and negative signals from mesoderm regulate the expression of mouse Otx2 in ectoderm explants. Development. 1994;120:2979–2989. PubMed

Bouillet P, Chazaud C, Oulad-Abdelghani M, Dolle P, Chambon P. Sequence and expression pattern of the Stra7 (Gbx-2) homeobox-containing gene induced by retinoic acid in P19 embryonal carcinoma cells. Dev Dyn. 1995;204:372–382. doi: 10.1002/aja.1002040404. PubMed DOI

Goto H, et al. Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation. J Biol Chem. 1999;274:25543–25549. doi: 10.1074/jbc.274.36.25543. PubMed DOI

Laurent A, Blasi F. Differential DNA damage signalling and apoptotic threshold correlate with mouse epiblast-specific hypersensitivity to radiation. Development. 2015;142:3675–3685. doi: 10.1242/dev.125708. PubMed DOI

Heyer BS, MacAuley A, Behrendtsen O, Werb Z. Hypersensitivity to DNA damage leads to increased apoptosis during early mouse development. Genes Dev. 2000;14:2072–2084. PubMed PMC

Fernandez-Diaz LC, et al. The absence of Prep1 causes p53-dependent apoptosis of mouse pluripotent epiblast cells. Development. 2010;137:3393–3403. doi: 10.1242/dev.050567. PubMed DOI

Guzman-Ayala M, et al. Chd1 is essential for the high transcriptional output and rapid growth of the mouse epiblast. Development. 2015;142:118–127. doi: 10.1242/dev.114843. PubMed DOI PMC

Panic L, et al. Ribosomal protein S6 gene haploinsufficiency is associated with activation of a p53-dependent checkpoint during gastrulation. Mol Cell Biol. 2006;26:8880–8891. doi: 10.1128/MCB.00751-06. PubMed DOI PMC

Ruland J, et al. p53 accumulation, defective cell proliferation, and early embryonic lethality in mice lacking tsg101. Proc Natl Acad Sci USA. 2001;98:1859–1864. doi: 10.1073/pnas.98.4.1859. PubMed DOI PMC

Singh AP, et al. Brg1 Enables Rapid Growth of the Early Embryo by Suppressing Genes That Regulate Apoptosis and Cell Growth Arrest. Mol Cell Biol. 2016;36:1990–2010. doi: 10.1128/MCB.01101-15. PubMed DOI PMC

Tarca AL, et al. A novel signaling pathway impact analysis. Bioinformatics. 2009;25:75–82. doi: 10.1093/bioinformatics/btn577. PubMed DOI PMC

Wang Q, et al. The p53 Family Coordinates Wnt and Nodal Inputs in Mesendodermal Differentiation of Embryonic Stem Cells. Cell Stem Cell. 2016 PubMed PMC

Bergmann JH, et al. Regulation of the ESC transcriptome by nuclear long noncoding RNAs. Genome Res. 2015;25:1336–1346. doi: 10.1101/gr.189027.114. PubMed DOI PMC

Collignon J, Varlet I, Robertson EJ. Relationship between asymmetric nodal expression and the direction of embryonic turning. Nature. 1996;381:155–158. doi: 10.1038/381155a0. PubMed DOI

Constam DB. Running the gauntlet: an overview of the modalities of travel employed by the putative morphogen Nodal. Curr Opin Genet Dev. 2009;19:302–307. doi: 10.1016/j.gde.2009.06.006. PubMed DOI

Constam DB. Riding shotgun: a dual role for the epidermal growth factor-Cripto/FRL-1/Cryptic protein Cripto in Nodal trafficking. Traffic. 2009;10:783–791. doi: 10.1111/j.1600-0854.2009.00874.x. PubMed DOI

Zhou X, Sasaki H, Lowe L, Hogan BL, Kuehn MR. Nodal is a novel TGF-beta-like gene expressed in the mouse node during gastrulation. Nature. 1993;361:543–547. doi: 10.1038/361543a0. PubMed DOI

Iratni R, et al. Inhibition of excess nodal signaling during mouse gastrulation by the transcriptional corepressor DRAP1. Science. 2002;298:1996–1999. doi: 10.1126/science.1073405. PubMed DOI

Meno C, et al. Mouse Lefty2 and zebrafish antivin are feedback inhibitors of nodal signaling during vertebrate gastrulation. Mol Cell. 1999;4:287–298. doi: 10.1016/S1097-2765(00)80331-7. PubMed DOI

Stuckey DW, et al. Coordination of cell proliferation and anterior-posterior axis establishment in the mouse embryo. Development. 2011;138:1521–1530. doi: 10.1242/dev.063537. PubMed DOI PMC

Brown EJ, Baltimore D. ATR disruption leads to chromosomal fragmentation and early embryonic lethality. Genes Dev. 2000;14:397–402. PubMed PMC

Dobles M, Liberal V, Scott ML, Benezra R, Sorger PK. Chromosome missegregation and apoptosis in mice lacking the mitotic checkpoint protein Mad2. Cell. 2000;101:635–645. doi: 10.1016/S0092-8674(00)80875-2. PubMed DOI

Hakem R, et al. The tumor suppressor gene Brca1 is required for embryonic cellular proliferation in the mouse. Cell. 1996;85:1009–1023. doi: 10.1016/S0092-8674(00)81302-1. PubMed DOI

Jeon Y, et al. TopBP1 deficiency causes an early embryonic lethality and induces cellular senescence in primary cells. J Biol Chem. 2011;286:5414–5422. doi: 10.1074/jbc.M110.189704. PubMed DOI PMC

Kalitsis P, Earle E, Fowler KJ, Choo KH. Bub3 gene disruption in mice reveals essential mitotic spindle checkpoint function during early embryogenesis. Genes Dev. 2000;14:2277–2282. doi: 10.1101/gad.827500. PubMed DOI PMC

Campbell PA, Perez-Iratxeta C, Andrade-Navarro MA, Rudnicki MA. Oct4 targets regulatory nodes to modulate stem cell function. PLoS One. 2007;2:e553. doi: 10.1371/journal.pone.0000553. PubMed DOI PMC

Guttman M, et al. lincRNAs act in the circuitry controlling pluripotency and differentiation. Nature. 2011;477:295–300. doi: 10.1038/nature10398. PubMed DOI PMC

Ivanova N, et al. Dissecting self-renewal in stem cells with RNA interference. Nature. 2006;442:533–538. doi: 10.1038/nature04915. PubMed DOI

Jacks T, et al. Tumor spectrum analysis in p53-mutant mice. Curr Biol. 1994;4:1–7. doi: 10.1016/S0960-9822(00)00002-6. PubMed DOI

Lawson, K. A. & Wilson, V. in Kaufman’s Atlas of Mouse Development Supplement: With Coronal Sections (eds R. Baldock, J. B. Bard, D. R. Davidson, & G. Morriss-Kay) Ch. 3, 51–64 (Elsevier, 2015).

Chuma S, Nakatsuji N. Autonomous transition into meiosis of mouse fetal germ cells in vitro and its inhibition by gp130-mediated signaling. Developmental biology. 2001;229:468–479. doi: 10.1006/dbio.2000.9989. PubMed DOI

Georgiades P, Rossant J. Ets2 is necessary in trophoblast for normal embryonic anteroposterior axis development. Development. 2006;133:1059–1068. doi: 10.1242/dev.02277. PubMed DOI

Polydorou C, Georgiades P. Ets2-dependent trophoblast signalling is required for gastrulation progression after primitive streak initiation. Nat Commun. 2013;4:1658. doi: 10.1038/ncomms2646. PubMed DOI

Carvalho BS, Irizarry RA. A framework for oligonucleotide microarray preprocessing. Bioinformatics. 2010;26:2363–2367. doi: 10.1093/bioinformatics/btq431. PubMed DOI PMC

Smyth, G. K. in Bioinforma. Comput. Biol. Solut. Using R Bioconductor (eds Gentleman, R. et al.) 397–420 (Springer, 2005).

Tian L, et al. Discovering statistically significant pathways in expression profiling studies. Proc Natl Acad Sci USA. 2005;102:13544–13549. doi: 10.1073/pnas.0506577102. PubMed DOI PMC

Maciejewski H. Gene set analysis methods: statistical models and methodological differences. Brief Bioinform. 2014;15:504–518. doi: 10.1093/bib/bbt002. PubMed DOI PMC