Regulation of histone H3 lysine 9 methylation in oocytes and early pre-implantation embryos

Epigenetic modifications of the genome, such as covalent modification of histone residues, ensure appropriate gene activation during pre-implantation development, and are probably involved in the asymmetric reprogramming of the parental genomes after fertilization. We investigated the methylation patterns of histone H3 at lysine 9 (H3/K9), and the regulatory mechanism involved in the asymmetric remodeling of parental genomes during early preimplantation development in mice. Immunocytochemistry with an antibody that specifically recognizes methylated H3/K9 showed a very weak or absent methylation signal in the male pronucleus, whereas a distinct methylation signal was detected in the female pronucleus. This asymmetric H3/K9 methylation pattern in the different parental genomes persisted until the two-cell stage. However, de novo methylation of H3/K9 occurred and the asymmetry was lost during the four-cell stage. The unmethylated male pronucleus underwent de novo methylation when it was transferred into enucleated GV- or MII-stage oocytes, which suggests that histone H3 methylase is active before fertilization, but not afterwards, and that the asymmetric methylation pattern is generated by this change in methylase activity in the cytoplasm after fertilization. Thus, histone H3 is methylated only in the maternal chromosomes, which are present in the oocytes before fertilization, and is not methylated in the paternal chromosomes, which are absent. The maintenance of asymmetric H3/K9 methylation patterns in early embryos is an active process that depends on protein synthesis and zygotic transcription, as de novo methylation in the male pronucleus occurred when either protein synthesis or gene expression was inhibited by cycloheximide orα -amanitin, respectively. In addition, corresponding de novo methylation of H3/K9 and DNA occurred when the male pronucleus was transferred to an enucleated GV oocyte. Our results suggest that H3/K9 methylation is an epigenetic marker of parental genome origin during early preimplantation development.

[1]  Jeannie T. Lee,et al.  Inheritance of a pre-inactivated paternal X chromosome in early mouse embryos , 2003, Nature.

[2]  E. Wolf,et al.  Epigenetic Marking Correlates with Developmental Potential in Cloned Bovine Preimplantation Embryos , 2003, Current Biology.

[3]  T. Jenuwein,et al.  Suv39h-Mediated Histone H3 Lysine 9 Methylation Directs DNA Methylation to Major Satellite Repeats at Pericentric Heterochromatin , 2003, Current Biology.

[4]  C. Allis,et al.  Trimethylated lysine 9 of histone H3 is a mark for DNA methylation in Neurospora crassa , 2003, Nature Genetics.

[5]  T. Mukai,et al.  Domain regulation of imprinting cluster in Kip2/Lit1 subdomain on mouse chromosome 7F4/F5: large-scale DNA methylation analysis reveals that DMR-Lit1 is a putative imprinting control region. , 2002, Genome research.

[6]  S. Jacobsen,et al.  DNA methylation controls histone H3 lysine 9 methylation and heterochromatin assembly in Arabidopsis , 2002, The EMBO journal.

[7]  B. Turner,et al.  Cellular Memory and the Histone Code , 2002, Cell.

[8]  Gioacchino Natoli,et al.  Dynamic changes in histone H3 Lys 9 methylation occurring at tightly regulated inducible inflammatory genes. , 2002, Genes & development.

[9]  F. Aoki,et al.  Analysis of the Mechanism for Chromatin Remodeling in Embryos Reconstructedby Somatic Nuclear Transfer1 , 2002, Biology of reproduction.

[10]  T. Bestor,et al.  Histone modification and replacement in chromatin activation. , 2002, Genes & development.

[11]  R. Martienssen,et al.  Dependence of Heterochromatic Histone H3 Methylation Patterns on the Arabidopsis Gene DDM1 , 2002, Science.

[12]  Tony Kouzarides,et al.  Histone methylation defines epigenetic asymmetry in the mouse zygote. , 2002, The International journal of developmental biology.

[13]  G. Maul,et al.  SETDB1: a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins. , 2002, Genes & development.

[14]  J. P. Jackson,et al.  Control of CpNpG DNA methylation by the KRYPTONITE histone H3 methyltransferase , 2002, Nature.

[15]  F. Aoki,et al.  Involvement of chromatin structure in the regulation of mouse zygotic gene activation , 2002 .

[16]  S. Elgin,et al.  Epigenetic Codes for Heterochromatin Formation and Silencing Rounding up the Usual Suspects , 2002, Cell.

[17]  D. Gilbert,et al.  Heterochromatin, HP1 and methylation at lysine 9 of histone H3 in animals , 2002, Chromosoma.

[18]  K. Mochida,et al.  Faithful expression of imprinted genes in cloned mice. , 2002, Science.

[19]  C. Allis,et al.  Methylation of Histone H3 at Lys-9 Is an Early Mark on the X Chromosome during X Inactivation , 2001, Cell.

[20]  E. Selker,et al.  A histone H3 methyltransferase controls DNA methylation in Neurospora crassa , 2001, Nature.

[21]  F. Aoki,et al.  Posttranscriptional Regulation of Cyclin A1 and Cyclin A2 During Mouse Oocyte Meiotic Maturation and Preimplantation Development1 , 2001, Biology of reproduction.

[22]  H. Worman,et al.  Transcriptional repression of euchromatic genes by Drosophila heterochromatin protein 1 and histone modifiers , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Sean D. Taverna,et al.  Specificity of the HP1 chromo domain for the methylated N‐terminus of histone H3 , 2001, The EMBO journal.

[24]  C. Allis,et al.  Translating the Histone Code , 2001, Science.

[25]  W. Reik,et al.  Epigenetic Reprogramming in Mammalian Development , 2001, Science.

[26]  M. Surani,et al.  Imprinting and the Epigenetic Asymmetry Between Parental Genomes , 2001, Science.

[27]  Ken-ichi Noma,et al.  Transitions in Distinct Histone H3 Methylation Patterns at the Heterochromatin Domain Boundaries , 2001, Science.

[28]  Andrew J. Bannister,et al.  Rb targets histone H3 methylation and HP1 to promoters , 2001, Nature.

[29]  J. Bender,et al.  Arabidopsis cmt3 chromomethylase mutations block non-CG methylation and silencing of an endogenous gene. , 2001, Genes & development.

[30]  B. Pickard,et al.  Epigenetic targeting in the mouse zygote marks DNA for later methylation: a mechanism for maternal effects in development , 2001, Mechanisms of Development.

[31]  Andrew J. Bannister,et al.  Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain , 2001, Nature.

[32]  J. Walter,et al.  Maternal methylation imprints on human chromosome 15 are established during or after fertilization , 2001, Nature Genetics.

[33]  D. Solter,et al.  Timely translation during the mouse oocyte-to-embryo transition. , 2000, Development.

[34]  R. Schultz,et al.  Histone macroH2A1 is concentrated in the inactive X chromosome of female preimplantation mouse embryos. , 2000, Development.

[35]  T. Haaf,et al.  Spatial Separation of Parental Genomes in Preimplantation Mouse Embryos , 2000, The Journal of cell biology.

[36]  J. Walter,et al.  Embryogenesis: Demethylation of the zygotic paternal genome , 2000, Nature.

[37]  R. Schultz,et al.  DNA replication in the 1-cell mouse embryo: stimulatory effect of histone acetylation , 1999, Zygote.

[38]  J. Renard,et al.  Differential H4 acetylation of paternal and maternal chromatin precedes DNA replication and differential transcriptional activity in pronuclei of 1-cell mouse embryos. , 1997, Development.

[39]  Z. Xu,et al.  Spontaneous activation of ovulated mouse eggs: time-dependent effects on M-phase exit, cortical granule exocytosis, maternal messenger ribonucleic acid recruitment, and inositol 1,4,5-trisphosphate sensitivity. , 1997, Biology of reproduction.

[40]  M. DePamphilis,et al.  Developmental acquisition of enhancer function requires a unique coactivator activity , 1997, The EMBO journal.

[41]  F. Aoki,et al.  Regulation of transcriptional activity during the first and second cell cycles in the preimplantation mouse embryo. , 1997, Developmental biology.

[42]  R. Kobayashi,et al.  Nucleosome Assembly by a Complex of CAF-1 and Acetylated Histones H3/H4 , 1996, Cell.

[43]  P. Debey,et al.  Endogenous transcription occurs at the 1-cell stage in the mouse embryo. , 1995, Experimental cell research.

[44]  C. Allis,et al.  Conservation of deposition-related acetylation sites in newly synthesized histones H3 and H4. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[45]  M. DePamphilis,et al.  Requirements for promoter activity in mouse oocytes and embryos distinguish paternal pronuclei from maternal and zygotic nuclei. , 1993, Developmental biology.

[46]  R. Schultz.  Regulation of zygotic gene activation in the mouse , 1993, BioEssays : news and reviews in molecular, cellular and developmental biology.

[47]  V. Chapman,et al.  Parental imprinting studied by allele-specific primer extension after PCR: paternal X chromosome-linked genes are transcribed prior to preferential paternal X chromosome inactivation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[48]  D L Spector,et al.  Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific alpha-satellite DNA sequences , 1992, The Journal of cell biology.

[49]  R. Schultz,et al.  Regulation of hsp70 mRNA levels during oocyte maturation and zygotic gene activation in the mouse. , 1991, Developmental biology.

[50]  J. Lewis,et al.  Development of 1-cell embryos from different strains of mice in CZB medium. , 1990, Biology of reproduction.

[51]  M. Lyon,et al.  Clonal analysis of X-chromosome inactivation and the origin of the germ line in the mouse embryo. , 1985, Journal of embryology and experimental morphology.

[52]  P. Braude,et al.  The transition from maternal to embryonic control in the 2‐cell mouse embryo. , 1982, The EMBO journal.

[53]  M. Sasaki,et al.  Preferential inactivation of the paternally derived X chromosome in the extraembryonic membranes of the mouse , 1975, Nature.

[54]  M. Lyon.  Gene Action in the X-chromosome of the Mouse (Mus musculus L.) , 1961, Nature.

[55]  T. Haaf.  The battle of the sexes after fertilization: behaviour of paternal and maternal chromosomes in the early mammalian embryo , 2004, Chromosome Research.

[56]  T. Mukai,et al.  Domain regulation of imprinting cluster in Kip2/Lit1 subdomain on mouse chromosome 7F4/F5: large-scale DNA methylation analysis reveals that DMR-Lit1 is a putative imprinting control region. , 2004, Genome research.

[57]  N. Brockdorff,et al.  Histone H3 lysine 9 methylation is an epigenetic imprint of facultative heterochromatin , 2002, Nature Genetics.

[58]  Wendy Dean,et al.  Dynamic reprogramming of DNA methylation in the early mouse embryo. , 2002, Developmental biology.

[59]  A. C. Chinault,et al.  Differentially methylated forms of histone H3 show unique association patterns with inactive human X chromosomes , 2002, Nature Genetics.

[60]  Kenneth Y. Tsai,et al.  Control of CpNpG DNA methylation by the KRYPTONITE histone H 3 methyltransferase , 2002 .

[61]  W. Reik,et al.  Genomic imprinting: parental influence on the genome , 2001, Nature Reviews Genetics.

[62]  A. Wolffe.  Chapter Three – Chromatin and Nuclear Assembly , 2000 .

[63]  J. Biggers,et al.  [9] Culture of preimplantation embryos , 1993 .

[64]  D. Spector,et al.  Macromolecular domains within the cell nucleus. , 1993, Annual review of cell biology.

[65]  J. Biggers,et al.  Culture of preimplantation embryos. , 1993, Methods in enzymology.

[66]  N. Wake,et al.  Cytologic evidence for preferential inactivation of the paternally derived X chromosome in XX mouse blastocysts. , 1978, Cytogenetics and cell genetics.