Skip to main content
Article thumbnail
Location of Repository

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

By Dario Nicetto, Matthias Hahn, Julia Jung, Tobias D. Schneider, Tobias Straub, Robert David, Gunnar Schotta and Ralph A. W. Rupp


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.

Topics: Medizin, ddc:610
Publisher: Ludwig-Maximilians-Universität München
Year: 2013
DOI identifier: 10.1371/journal.pgen.1003188
OAI identifier:
Provided by: Open Access LMU

Suggested articles


  1. (2008). A chromatin-wide transition to H4K20 monomethylation impairs genome integrity and programmed DNA rearrangements in the mouse.
  2. (2009). A hierarchy of H3K4me3 and H3K27me3 acquisition in spatial gene regulation in Xenopus embryos.
  3. (2004). A silencing pathway to induce H3-K9 and H4-K20 trimethylation at constitutive heterochromatin.
  4. (2009). Chromatin immunoprecipitation in early Xenopus laevis embryos.
  5. (2006). Chromatin in pluripotent embryonic stem cells and differentiation.
  6. (2010). Chromatin signature of embryonic pluripotency is established during genome activation.
  7. (2006). Conserved roles for Oct4 homologues in maintaining multipotency during early vertebrate development.
  8. (2010). de la Serna IL
  9. (2004). Distinct dynamics and distribution of histone methyl-lysine derivatives in mouse development.
  10. (2010). Epigenetic control of embryonic stem cell fate.
  11. (2009). Epigenetic dynamics of stem cells and cell lineage commitment: digging Waddington’s canal.
  12. (2010). Epigenetic regulation of development by histone lysine methylation.
  13. (2002). G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis.
  14. (2004). gene and counteracts terminal differentiation in Xenopus embryos.
  15. (2007). Genomewide maps of chromatin state in pluripotent and lineage-committed cells.
  16. (2007). Highresolution profiling of histone methylations in the human genome.
  17. (2004). Histone deacetylase activity is required for embryonic stem cell differentiation.
  18. (2012). Histone h1 depletion impairs embryonic stem cell differentiation.
  19. (2011). Histone H4 Lys 20 monomethylation by histone methylase SET8 mediates Wnt target gene activation.
  20. (2010). Histone variants in metazoan development.
  21. (2007). How is pluripotency determined and maintained?
  22. (2006). Hyperdynamic plasticity of chromatin proteins in pluripotent embryonic stem cells.
  23. LeeBM,MahadevanLC(2009)Stabilityofhistonemodificationsacrossmammalian genomes: implications for ‘epigenetic’ marking.
  24. MagklaraA,YenA,ColquittBM,Clowney EJ,AllenW,etal.(2011) Anepigenetic signature for monoallelic olfactory receptor expression.
  25. (2003). Mechanism of histone lysine methyl transfer revealed by the structure of SET7/9-AdoMet.
  26. (2008). MesP1 drives vertebrate cardiovascular differentiation through Dkk-1-mediated blockade of Wnt-signalling.
  27. (2002). Molecular cloning of ESET, a novel histone H3-specific methyltransferase that interacts with ERG transcription factor.
  28. (2009). Neural induction and factors that stabilize a neural fate. Birth defects research Part C, Embryo today : reviews 87:
  29. (2002). Neural induction, the default model and embryonic stem cells.
  30. (1991). Neuronal determination without cell division in Xenopus embryos.
  31. (2008). Oct25 represses transcription of nodal/activin target genes by interaction with signal transducers during Xenopus gastrulation.
  32. (2011). Pluripotency factors in embryonic stem cells regulate differentiation into germ layers.
  33. (2011). Pluripotency factors regulate definitive endoderm specification through eomesodermin.
  34. (2006). Polycomb complexes repress developmental regulators in murine embryonic stem cells.
  35. (2000). Regulation of chromatin structure by site-specific histone H3 methyltransferases.
  36. (1992). Sequential expression of multiple POU proteins during amphibian early development.
  37. (2007). Shilatifard A
  38. (2011). Stage-specific histone modification profiles reveal global transitions in the Xenopus embryonic epigenome.
  39. (2007). Suzuki A
  40. (2004). The functions of E(Z)/EZH2-mediated methylation of lysine 27 in histone H3.
  41. (2010). The genome of the Western clawed frog Xenopus tropicalis.
  42. (2003). The role of DNA methylation in setting up chromatin structure during development.
  43. (2005). The SET-domain protein superfamily: protein lysine methyltransferases.
  44. Vertino PM (2011) SUV420H2-mediated H4K20 trimethylation enforces RNA polymerase II promoter-proximal pausing by blocking hMOF-dependent H4K16 acetylation.
  45. (2006). Xenopus laevis POU91 protein, an Oct3/4 homologue, regulates competence transitions from mesoderm to neural cell fates.
  46. (2006). Xenopus POU factors of subclass V inhibit activin/nodal signaling during gastrulation.
  47. (1996). Xenopus Xsal-1, a vertebrate homolog of the region specific homeotic gene spalt of Drosophila.

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.