RESUMEN
Genomic imprinting in mammals is controlled by DNA methylation imprints that are acquired in the gametes, at essential sequence elements called 'imprinting control regions' (ICRs). What signals paternal imprint acquisition in male germ cells remains unknown. To address this question, we explored histone methylation at ICRs in mouse primordial germ cells (PGCs). By 13.5 days post coitum (d.p.c.), H3 lysine-9 and H4 lysine-20 trimethylation are depleted from ICRs in male (and female) PGCs, indicating that these modifications do not signal subsequent imprint acquisition, which initiates at â¼15.5 d.p.c. Furthermore, during male PGC development, H3 lysine-4 trimethylation becomes biallelically enriched at 'maternal' ICRs, which are protected against DNA methylation, and whose promoters are active in the male germ cells. Remarkably, high transcriptional read-through is detected at the paternal ICRs H19-DMR and Ig-DMR at the time of imprint establishment, from one of the strands predominantly. Combined, our data evoke a model in which differential histone modification states linked to transcriptional events may signal the specificity of imprint acquisition during spermatogenesis.
Asunto(s)
Metilación de ADN , Impresión Genómica , Histonas/metabolismo , Transcripción Genética , Animales , Separación Celular , Inmunoprecipitación de Cromatina , Drosophila melanogaster , Femenino , Citometría de Flujo , MasculinoRESUMEN
In the mid-1980s, elegant studies on mouse embryos revealed that both parental genomes are required for normal development leading to the discovery of genomic imprinting. Imprinting is a parent-of-origin-dependent epigenetic mechanism whereby a subset of autosomal genes is expressed from only one of the parental alleles. Imprinting control involves both DNA- and histone-methylation, which differentially mark the parental alleles. More than a hundred imprinted genes have been identified so far, many of which play important roles in the regulation of growth and development. Nonetheless, the full extent of imprinting and its biological functions remain underestimated. In this review, we describe recently developed strategies to identify novel imprinted genes and highlight the potential of combining several high throughput approaches. By integrating databases obtained from epigenome- and transcriptome-wide analyses, we now have the unique opportunity to identify all the imprinted genes in the human/mouse genomes.
Asunto(s)
Impresión Genómica/genética , Alelos , Animales , Cromatina/genética , Epigénesis Genética/genética , Humanos , Modelos Genéticos , Transcripción Genética/genéticaRESUMEN
Mono-allelic expression of imprinted genes from either the paternal or the maternal allele is mediated by imprinting control regions (ICRs), which are epigenetically marked in an allele-specific fashion. Although, in somatic cells, these epigenetic marks comprise both DNA methylation and histone methylation, the relationship between these two modifications in imprint acquisition and maintenance remains unclear. To address this important question, we analyzed histone modifications at ICRs in mid-gestation embryos that were obtained from Dnmt3L(-/-) females, in which DNA methylation imprints at ICRs are not established during oogenesis. The absence of maternal DNA methylation imprints in these conceptuses led to a marked decrease and loss of allele-specificity of the repressive H3K9me3, H4K20me3 and H2A/H4R3me2 histone modifications, providing the first evidence of a mechanistic link between DNA and histone methylation at ICRs. The existence of this relationship was strengthened by the observation that when DNA methylation was still present at the Snrpn and Peg3 ICRs in some of the progeny of Dnmt3L(-/-) females, these ICRs were associated with the usual patterns of histone methylation. Combined, our data establish that DNA methylation is involved in the acquisition and/or maintenance of histone methylation at ICRs.
Asunto(s)
Metilación de ADN , Genoma , Impresión Genómica , Histonas/metabolismo , Alelos , Animales , ADN (Citosina-5-)-Metiltransferasas/deficiencia , ADN (Citosina-5-)-Metiltransferasas/genética , ADN (Citosina-5-)-Metiltransferasas/metabolismo , Embrión de Mamíferos/metabolismo , Femenino , Proteína Adaptadora GRB10/genética , Proteína Adaptadora GRB10/metabolismo , Masculino , Metilación , Ratones , Ratones Noqueados , Activación Transcripcional , Proteínas Nucleares snRNP/genética , Proteínas Nucleares snRNP/metabolismoRESUMEN
At the time of fertilisation, the parental genomes have a strikingly different organisation. Sperm DNA is packaged globally with protamines, whereas the oocyte's genome is wrapped around nucleosomes. The maternal and paternal genomes are functionally different as well, and are both required for normal uterine and postnatal development. The functional requirement of both parental genomes is a consequence of differential epigenetic marking by DNA methylation during oogenesis and spermatogenesis, on a group of genes called imprinted genes. After fertilisation, these parental marks persist throughout development and convey the allelic expression of imprinted genes. Pathological perturbation of methylation imprints, before fertilisation in the germ cells, or during development, gives rise to growth-related syndromes, and is frequently observed in cancer as well. Alteration of imprints is thought to occur early in carcinogenesis and shows similarities with the acquisition of aberrant DNA methylation at tumour suppressor genes. This suggests that similar underlying mechanisms could be involved.
Asunto(s)
Epigénesis Genética , Neoplasias/genética , Metilación de ADN/genética , Femenino , Desarrollo Fetal/genética , Terapia Genética/métodos , Humanos , Masculino , Neoplasias/terapia , Oligospermia/genética , PadresRESUMEN
Genomic imprinting is a developmental mechanism that mediates parent-of-origin-specific expression in a subset of genes. How the tissue specificity of imprinted gene expression is controlled remains poorly understood. As a model to address this question, we studied Grb10, a gene that displays brain-specific expression from the paternal chromosome. Here, we show in the mouse that the paternal promoter region is marked by allelic bivalent chromatin enriched in both H3K4me2 and H3K27me3, from early embryonic stages onwards. This is maintained in all somatic tissues, but brain. The bivalent domain is resolved upon neural commitment, during the developmental window in which paternal expression is activated. Our data indicate that bivalent chromatin, in combination with neuronal factors, controls the paternal expression of Grb10 in brain. This finding highlights a novel mechanism to control tissue-specific imprinting.
Asunto(s)
Alelos , Cromatina/metabolismo , Proteína Adaptadora GRB10 , Impresión Genómica , Animales , Encéfalo/citología , Encéfalo/embriología , Encéfalo/fisiología , Diferenciación Celular , Células Cultivadas , Cromatina/genética , Islas de CpG , Embrión de Mamíferos/anatomía & histología , Embrión de Mamíferos/fisiología , Femenino , Proteína Adaptadora GRB10/genética , Proteína Adaptadora GRB10/metabolismo , Histonas/genética , Histonas/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuronas/citología , Neuronas/fisiología , Complejo Represivo Polycomb 2 , Regiones Promotoras Genéticas , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Células Madre/fisiologíaRESUMEN
Whereas DNA methylation is essential for genomic imprinting, the importance of histone methylation in the allelic expression of imprinted genes is unclear. Imprinting control regions (ICRs), however, are marked by histone H3-K9 methylation on their DNA-methylated allele. In the placenta, the paternal silencing along the Kcnq1 domain on distal chromosome 7 also correlates with the presence of H3-K9 methylation, but imprinted repression at these genes is maintained independently of DNA methylation. To explore which histone methyltransferase (HMT) could mediate the allelic H3-K9 methylation on distal chromosome 7, and at ICRs, we generated mouse conceptuses deficient for the SET domain protein G9a. We found that in the embryo and placenta, the differential DNA methylation at ICRs and imprinted genes is maintained in the absence of G9a. Accordingly, in embryos, imprinted gene expression was unchanged at the domains analyzed, in spite of a global loss of H3-K9 dimethylation (H3K9me2). In contrast, the placenta-specific imprinting of genes on distal chromosome 7 is impaired in the absence of G9a, and this correlates with reduced levels of H3K9me2 and H3K9me3. These findings provide the first evidence for the involvement of an HMT and suggest that histone methylation contributes to imprinted gene repression in the trophoblast.
Asunto(s)
Impresión Genómica , N-Metiltransferasa de Histona-Lisina/fisiología , Histonas/metabolismo , Placenta/enzimología , Animales , Embrión de Mamíferos , Femenino , Silenciador del Gen , Histona Metiltransferasas , Metilación , Ratones , Proteína Metiltransferasas , Trofoblastos/metabolismoRESUMEN
It is generally assumed that the developmental program of embryogenesis relies on epigenetic mechanisms. However, a mechanistic link between epigenetic marks and cell fate decisions had not been established so far. In a recent article, Torres-Padilla and colleagues show that epigenetic information and, more precisely, histone arginine methylation mediated by CARM1 could contribute to cell fate decisions in the mouse 4-cell-stage embryo. It provides the first indications that global epigenetic information influences allocation of pluripotent cells toward the first cell lineages.