Suv4-20h Histone Methyltransferases Promote Neuroectodermal Differentiation by Silencing the Pluripotency-Associated Oct-25 Gene

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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.

Vyšlo v časopise: Suv4-20h Histone Methyltransferases Promote Neuroectodermal Differentiation by Silencing the Pluripotency-Associated Oct-25 Gene. PLoS Genet 9(1): e32767. doi:10.1371/journal.pgen.1003188
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003188

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Zdroje

1. NiwaH (2007) How is pluripotency determined and maintained? Development 134: 635–646.

2. MeshorerE, YellajoshulaD, GeorgeE, ScamblerPJ, BrownDT, et al. (2006) Hyperdynamic plasticity of chromatin proteins in pluripotent embryonic stem cells. Dev Cell 10: 105–116.

3. HembergerM, DeanW, ReikW (2009) Epigenetic dynamics of stem cells and cell lineage commitment: digging Waddington's canal. Nat Rev Mol Cell Bio 10: 526–537.

4. SaladiSV, de la SernaIL (2010) ATP dependent chromatin remodeling enzymes in embryonic stem cells. Stem Cell Rev 6: 62–73.

5. SinghalN, GraumannJ, WuG, Arauzo-BravoMJ, HanDW, et al. (2010) Chromatin-Remodeling Components of the BAF Complex Facilitate Reprogramming. Cell 141: 943–955.

6. BanaszynskiLA, AllisCD, LewisPW (2010) Histone variants in metazoan development. Dev Cell 19: 662–674.

7. LeeBM, MahadevanLC (2009) Stability of histone modifications across mammalian genomes: implications for ‘epigenetic’ marking. J Cell Biochem 108: 22–34.

8. MeshorerE, MisteliT (2006) Chromatin in pluripotent embryonic stem cells and differentiation. Nat Rev Mol Cell Bio 7: 540–546.

9. BironVL, McManusKJ, HuN, HendzelMJ, UnderhillDA (2004) Distinct dynamics and distribution of histone methyl-lysine derivatives in mouse development. Dev Biol 276: 337–351.

10. HashimshonyT, ZhangJ, KeshetI, BustinM, CedarH (2003) The role of DNA methylation in setting up chromatin structure during development. Nat Genet 34: 187–192.

11. LeeJH, HartSR, SkalnikDG (2004) Histone deacetylase activity is required for embryonic stem cell differentiation. Genesis 38: 32–38.

12. AkkersRC, van HeeringenSJ, JacobiUG, Janssen-MegensEM, FrancoijsKJ, et al. (2009) A hierarchy of H3K4me3 and H3K27me3 acquisition in spatial gene regulation in Xenopus embryos. Dev Cell 17: 425–434.

13. SchneiderTD, Arteaga-SalasJM, MenteleE, DavidR, NicettoD, et al. (2011) Stage-specific histone modification profiles reveal global transitions in the Xenopus embryonic epigenome. PLoS ONE 6: e22548 doi:10.1371/journal.pone.0022548.

14. VastenhouwNL, ZhangY, WoodsIG, ImamF, RegevA, et al. (2010) Chromatin signature of embryonic pluripotency is established during genome activation. Nature 464: 922–926.

15. BhaumikSR, SmithE, ShilatifardA (2007) Covalent modifications of histones during development and disease pathogenesis. Nat Struct Mol Biol 14: 1008–1016.

16. DambacherS, HahnM, SchottaG (2010) Epigenetic regulation of development by histone lysine methylation. Heredity 105: 24–37.

17. BoyerLA, PlathK, ZeitlingerJ, BrambrinkT, MedeirosLA, et al. (2006) Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature 441: 349–353.

18. CaoR, ZhangY (2004) The functions of E(Z)/EZH2-mediated methylation of lysine 27 in histone H3. Curr Opin Genet Dev 14: 155–164.

19. ChristophersenNS, HelinK (2010) Epigenetic control of embryonic stem cell fate. J Exp Med 207: 2287–2295.

20. ReaS, EisenhaberF, O'CarrollD, StrahlBD, SunZW, et al. (2000) Regulation of chromatin structure by site-specific histone H3 methyltransferases. Nature 406: 593–599.

21. SchottaG, LachnerM, SarmaK, EbertA, SenguptaR, et al. (2004) A silencing pathway to induce H3-K9 and H4-K20 trimethylation at constitutive heterochromatin. Gene Dev 18: 1251–1262.

22. TachibanaM, SugimotoK, NozakiM, UedaJ, OhtaT, et al. (2002) G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis. Gene Dev 16: 1779–1791.

23. YangL, XiaL, WuDY, WangH, ChanskyHA, et al. (2002) Molecular cloning of ESET, a novel histone H3-specific methyltransferase that interacts with ERG transcription factor. Oncogene 21: 148–152.

24. SchottaG, SenguptaR, KubicekS, MalinS, KauerM, et al. (2008) A chromatin-wide transition to H4K20 monomethylation impairs genome integrity and programmed DNA rearrangements in the mouse. Gene Dev 22: 2048–2061.

25. DillonSC, ZhangX, TrievelRC, ChengX (2005) The SET-domain protein superfamily: protein lysine methyltransferases. Genome Biol 6: 227.

26. KwonT, ChangJH, KwakE, LeeCW, JoachimiakA, et al. (2003) Mechanism of histone lysine methyl transfer revealed by the structure of SET7/9-AdoMet. EMBO J 22: 292–303.

27. RogersCD, MoodySA, CaseyES (2009) Neural induction and factors that stabilize a neural fate. Birth defects research Part C, Embryo today : reviews 87: 249–262.

28. HarrisWA, HartensteinV (1991) Neuronal determination without cell division in Xenopus embryos. Neuron 6: 499–515.

29. HinkleyCS, MartinJF, LeibhamD, PerryM (1992) Sequential expression of multiple POU proteins during amphibian early development. Mol Cell Biol 12: 638–649.

30. MorrisonGM, BrickmanJM (2006) Conserved roles for Oct4 homologues in maintaining multipotency during early vertebrate development. Development 133: 2011–2022.

31. CaoY, KnochelS, DonowC, MietheJ, KaufmannE, et al. (2004) The POU factor Oct-25 regulates the Xvent-2B gene and counteracts terminal differentiation in Xenopus embryos. J Biol Chem 279: 43735–43743.

32. CaoY, SiegelD, OswaldF, KnochelW (2008) Oct25 represses transcription of nodal/activin target genes by interaction with signal transducers during Xenopus gastrulation. J Biol Chem 283: 34168–34177.

33. SnirM, OfirR, EliasS, FrankD (2006) Xenopus laevis POU91 protein, an Oct3/4 homologue, regulates competence transitions from mesoderm to neural cell fates. EMBO J 25: 3664–3674.

34. Takebayashi-SuzukiK, AritaN, MurasakiE, SuzukiA (2007) The Xenopus POU class V transcription factor XOct-25 inhibits ectodermal competence to respond to bone morphogenetic protein-mediated embryonic induction. Mech Develop 124: 840–855.

35. HellstenU, HarlandRM, GilchristMJ, HendrixD, JurkaJ, et al. (2010) The genome of the Western clawed frog Xenopus tropicalis. Science 328: 633–636.

36. BarskiA, CuddapahS, CuiK, RohTY, SchonesDE, et al. (2007) High-resolution profiling of histone methylations in the human genome. Cell 129: 823–837.

37. MikkelsenTS, KuM, JaffeDB, IssacB, LiebermanE, et al. (2007) Genome-wide maps of chromatin state in pluripotent and lineage-committed cells. Nature 448: 553–560.

38. BoyerLA, LeeTI, ColeMF, JohnstoneSE, LevineSS, et al. (2005) Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122: 947–956.

39. Munoz-SanjuanI, BrivanlouAH (2002) Neural induction, the default model and embryonic stem cells. Nat Rev Neurosci 3: 271–280.

40. CaoY, SiegelD, KnochelW (2006) Xenopus POU factors of subclass V inhibit activin/nodal signaling during gastrulation. Mech Develop 123: 614–625.

41. TeoAK, ArnoldSJ, TrotterMW, BrownS, AngLT, et al. (2011) Pluripotency factors regulate definitive endoderm specification through eomesodermin. Gene Dev 25: 238–250.

42. ThomsonM, LiuSJ, ZouLN, SmithZ, MeissnerA, et al. (2011) Pluripotency factors in embryonic stem cells regulate differentiation into germ layers. Cell 145: 875–889.

43. DavidR, BrennerC, StieberJ, SchwarzF, BrunnerS, et al. (2008) MesP1 drives vertebrate cardiovascular differentiation through Dkk-1-mediated blockade of Wnt-signalling. Nature Cell Biol 10: 338–345.

44. Kapoor-VaziraniP, KageyJD, VertinoPM (2011) SUV420H2-mediated H4K20 trimethylation enforces RNA polymerase II promoter-proximal pausing by blocking hMOF-dependent H4K16 acetylation. Mol Cell Biol 31: 1594–1609.

45. MagklaraA, YenA, ColquittBM, ClowneyEJ, AllenW, et al. (2011) An epigenetic signature for monoallelic olfactory receptor expression. Cell 145: 555–570.

46. ZhangY, CookeM, PanjwaniS, CaoK, KrauthB, et al. (2012) Histone h1 depletion impairs embryonic stem cell differentiation. PLoS Genet 8: e1002691 doi:10.1371/journal.pgen.1002691.

47. LiZ, NieF, WangS, LiL (2011) Histone H4 Lys 20 monomethylation by histone methylase SET8 mediates Wnt target gene activation. P Natl Acad Sci USA 108: 3116–3123.

48. HollemannT, SchuhR, PielerT, StickR (1996) Xenopus Xsal-1, a vertebrate homolog of the region specific homeotic gene spalt of Drosophila. Mech of Develop 55: 19–32.

49. BlytheSA, ReidCD, KesslerDS, KleinPS (2009) Chromatin immunoprecipitation in early Xenopus laevis embryos. Dev Dynam 238: 1422–1432.