Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 11 de 11
Filter
Add more filters










Publication year range
1.
Proc Natl Acad Sci U S A ; 115(41): 10387-10391, 2018 10 09.
Article in English | MEDLINE | ID: mdl-30257947

ABSTRACT

Following erasure in the blastocyst, the entire genome undergoes de novo methylation at the time of implantation, with CpG islands being protected from this process. This bimodal pattern is then preserved throughout development and the lifetime of the organism. Using mouse embryonic stem cells as a model system, we demonstrate that the binding of an RNA polymerase complex on DNA before de novo methylation is predictive of it being protected from this modification, and tethering experiments demonstrate that the presence of this complex is, in fact, sufficient to prevent methylation at these sites. This protection is most likely mediated by the recruitment of enzyme complexes that methylate histone H3K4 over a local region and, in this way, prevent access to the de novo methylation complex. The topological pattern of H3K4me3 that is formed while the DNA is as yet unmethylated provides a strikingly accurate template for modeling the genome-wide basal methylation pattern of the organism. These results have far-reaching consequences for understanding the relationship between RNA transcription and DNA methylation.


Subject(s)
Blastocyst Inner Cell Mass/metabolism , DNA Methylation , Embryo, Mammalian/metabolism , Gene Expression Regulation, Developmental , Histones/metabolism , Transcription, Genetic , Animals , Blastocyst Inner Cell Mass/cytology , CpG Islands , DNA-Directed RNA Polymerases/metabolism , Embryo, Mammalian/cytology , Mice , Mice, Transgenic , Transcription Factors/metabolism
2.
Int J Dev Biol ; 61(3-4-5): 285-292, 2017.
Article in English | MEDLINE | ID: mdl-28621425

ABSTRACT

Fragile X syndrome is the most frequent cause of inherited intellectual disability. The primary molecular defect in this disease is the expansion of a CGG repeat in the 5' region of the fragile X mental retardation1 (FMR1) gene, leading to de novo methylation of the promoter and inactivation of this otherwise normal gene, but little is known about how these epigenetic changes occur during development. In order to gain insight into the nature of this process, we have used cell fusion technology to recapitulate the events that occur during early embryogenesis. These experiments suggest that the naturally occurring Fragile XFMR1 5' region undergoes inactivation post implantation in a Dicer/Ago-dependent targeted process which involves local SUV39H-mediated tri-methylation of histone H3K9. It thus appears that Fragile X syndrome may come about through inadvertent siRNA-mediated heterochromatinization.


Subject(s)
DNA Methylation , Epigenesis, Genetic , Fragile X Mental Retardation Protein/genetics , Fragile X Syndrome/genetics , Gene Expression Regulation, Developmental , 5' Untranslated Regions , Animals , Cell Differentiation , Embryonic Development , Embryonic Stem Cells/metabolism , Fibroblasts/metabolism , Heterochromatin/chemistry , Histones/metabolism , Humans , Mice , Nerve Tissue Proteins/genetics , Phenotype , Promoter Regions, Genetic , RNA/metabolism , RNA Interference , RNA, Small Interfering/metabolism
3.
Genes Dev ; 29(9): 923-33, 2015 May 01.
Article in English | MEDLINE | ID: mdl-25934504

ABSTRACT

DNA methylation patterns are set up in a relatively fixed programmed manner during normal embryonic development and are then stably maintained. Using genome-wide analysis, we discovered a postnatal pathway involving gender-specific demethylation that occurs exclusively in the male liver. This demodification is programmed to take place at tissue-specific enhancer sequences, and our data show that the methylation state at these loci is associated with and appears to play a role in the transcriptional regulation of nearby genes. This process is mediated by the secretion of testosterone at the time of sexual maturity, but the resulting methylation profile is stable and therefore can serve as an epigenetic memory even in the absence of this inducer. These findings add a new dimension to our understanding of the role of DNA methylation in vivo and provide the foundations for deciphering how environment can impact on the epigenetic regulation of genes in general.


Subject(s)
DNA Methylation , Epigenesis, Genetic/genetics , Liver/metabolism , Androgens/pharmacology , Animals , Castration , DNA Methylation/drug effects , Enhancer Elements, Genetic/genetics , Epigenesis, Genetic/drug effects , Female , Gene Expression Regulation, Developmental , Genome-Wide Association Study , Histones/genetics , Histones/metabolism , Humans , Liver/drug effects , Male , Mice , Mice, Inbred C57BL , Sex Characteristics , Testosterone/metabolism , Testosterone/pharmacology
4.
Nat Struct Mol Biol ; 21(1): 110-2, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24336222

ABSTRACT

After erasure in the early animal embryo, a new bimodal DNA methylation pattern is regenerated at implantation. We have identified a demethylation pathway in mouse embryonic cells that uses hydroxymethylation (Tet1), deamination (Aid), glycosylation (Mbd4) and excision repair (Gadd45a) genes. Surprisingly, this demethylation system is not necessary for generating the overall bimodal methylation pattern but does appear to be involved in resetting methylation patterns during somatic-cell reprogramming.


Subject(s)
DNA Methylation , Embryonic Stem Cells/metabolism , Amination , Animals , DNA Repair/genetics , Mice
5.
Int J Gynecol Cancer ; 21(2): 269-79, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21270610

ABSTRACT

BACKGROUND: DNA methylation regulates gene expression during development. The methylation pattern is established at the time of implantation. CpG islands are genome regions usually protected from methylation; however, selected islands are methylated later. Many undergo methylation in cancer, causing epigenetic gene silencing. Aberrant methylation occurs early in tumorigenesis, in a specific pattern, inhibiting differentiation.Although methylation of specific genes in ovarian tumors has been demonstrated in numerous studies, they represent only a fraction of all methylated genes in tumorigenesis. OBJECTIVES: To explore the hypermethylation design in ovarian cancer compared with the methylation profile of normal ovaries, on a genome-wide scale, thus shedding light on the role of gene silencing in ovarian carcinogenesis.Identifying genes that undergo de novo methylation in ovarian cancer may assist in creating biomarkers for disease diagnosis, prognosis, and treatment responsiveness. METHODS: DNA was collected from human epithelial ovarian cancers and normal ovaries. Methylation was detected by immunoprecipitation using 5-methyl-cytosine-antibodies. DNA was hybridized to a CpG island microarray containing 237,220 gene promoter probes. Results were analyzed by hybridization intensity, validated by bisulfite analysis. RESULTS: : A total of 367 CpG islands were specifically methylated in cancer cells. There was enrichment of methylated genes in functional categories related to cell differentiation and proliferation inhibition. It seems that their silencing enables tumor proliferation. CONCLUSIONS: This study provides new perspectives on methylation in ovarian carcinoma, genome-wide. It illustrates how methylation of CpG islands causes silencing of genes that have a role in cell differentiation and functioning. It creates potential biomarkers for diagnosis, prognosis, and treatment responsiveness.


Subject(s)
DNA Methylation , Neoplasms, Glandular and Epithelial , Ovarian Neoplasms , Carcinoma, Ovarian Epithelial , CpG Islands , Gene Silencing , Neoplasms, Glandular and Epithelial/genetics , Ovarian Neoplasms/genetics
6.
J Biol Chem ; 282(16): 12194-200, 2007 Apr 20.
Article in English | MEDLINE | ID: mdl-17311920

ABSTRACT

A large fraction of the animal genome is maintained in a transcriptionally repressed state throughout development. By generating viable Dnmt1(-)(/)(-) mouse cells we have been able to study the effect of DNA methylation on both gene expression and chromatin structure. Our results confirm that the underlying methylation pattern has a profound effect on histone acetylation and is the major effector of me-H3(K4) in the animal genome. We demonstrate that many methylated genes are subject to additional repression mechanisms that also impact on histone acetylation, and the data suggest that late replication timing may play an important role in this process.


Subject(s)
DNA (Cytosine-5-)-Methyltransferases/genetics , DNA (Cytosine-5-)-Methyltransferases/physiology , DNA Methylation , Gene Expression Regulation , Acetylation , Animals , Chromatin/chemistry , CpG Islands , DNA/chemistry , DNA (Cytosine-5-)-Methyltransferase 1 , DNA Replication , Epigenesis, Genetic , Fibroblasts/metabolism , Histones/chemistry , Mice , Mice, Transgenic , Oligonucleotide Array Sequence Analysis
7.
Nat Genet ; 39(2): 232-6, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17200670

ABSTRACT

Many genes associated with CpG islands undergo de novo methylation in cancer. Studies have suggested that the pattern of this modification may be partially determined by an instructive mechanism that recognizes specifically marked regions of the genome. Using chromatin immunoprecipitation analysis, here we show that genes methylated in cancer cells are specifically packaged with nucleosomes containing histone H3 trimethylated on Lys27. This chromatin mark is established on these unmethylated CpG island genes early in development and then maintained in differentiated cell types by the presence of an EZH2-containing Polycomb complex. In cancer cells, as opposed to normal cells, the presence of this complex brings about the recruitment of DNA methyl transferases, leading to de novo methylation. These results suggest that tumor-specific targeting of de novo methylation is pre-programmed by an established epigenetic system that normally has a role in marking embryonic genes for repression.


Subject(s)
DNA Methylation , Histones/metabolism , Neoplasms/genetics , Caco-2 Cells , Carrier Proteins , Cells, Cultured , Colonic Neoplasms/genetics , CpG Islands/genetics , Epigenesis, Genetic , Humans , Lysine/metabolism , Methylation , Methyltransferases/metabolism , Viral Envelope Proteins
8.
Nat Genet ; 38(2): 149-53, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16444255

ABSTRACT

DNA methylation has a role in the regulation of gene expression during normal mammalian development but can also mediate epigenetic silencing of CpG island genes in cancer and other diseases. Many individual genes (including tumor suppressors) have been shown to undergo de novo methylation in specific tumor types, but the biological logic inherent in this process is not understood. To decipher this mechanism, we have adopted a new approach for detecting CpG island DNA methylation that can be used together with microarray technology. Genome-wide analysis by this technique demonstrated that tumor-specific methylated genes belong to distinct functional categories, have common sequence motifs in their promoters and are found in clusters on chromosomes. In addition, many are already repressed in normal cells. These results are consistent with the hypothesis that cancer-related de novo methylation may come about through an instructive mechanism.


Subject(s)
DNA Methylation , Gene Expression Regulation, Neoplastic , Models, Genetic , Neoplasms/genetics , Animals , Chromosomes/genetics , Computational Biology , Genome , Neoplasms/pathology
9.
Curr Biol ; 14(11): 1007-12, 2004 Jun 08.
Article in English | MEDLINE | ID: mdl-15182675

ABSTRACT

The H19 imprinted gene locus is regulated by an upstream 2 kb imprinting control region (ICR) that influences allele-specific expression, DNA methylation, and replication timing. This ICR becomes de novo methylated during late spermatogenesis in the male but emerges from oogenesis in an unmethylated form, and this allele-specific pattern is then maintained throughout early development and in all tissues of the mouse. We have used a genetic approach involving transfection into embryonic stem (ES) cells in order to decipher how the maternal allele is protected from de novo methylation at the time of implantation. Our studies show that CCCTC binding factor (CTCF) boundary elements within the ICR have the ability to prevent de novo methylation on the maternal allele. Since CTCF does not recognize its binding sequence when methylated, this reaction does not occur on the paternal allele, thus preserving the gamete-derived, allele-specific pattern. These results suggest that CTCF may play a general role in the maintenance of differential methylation patterns in vivo.


Subject(s)
Alleles , DNA Methylation , DNA-Binding Proteins/metabolism , Genomic Imprinting/physiology , Locus Control Region/physiology , Repressor Proteins/metabolism , Animals , Base Sequence , Blotting, Southern , CCCTC-Binding Factor , Cells, Cultured , DNA-Binding Proteins/genetics , DNA-Binding Proteins/physiology , Female , Male , Mice , Mice, Transgenic , Plasmids/genetics , Repressor Proteins/genetics , Repressor Proteins/physiology , Sex Factors , Stem Cells/physiology , Transfection
10.
Nat Genet ; 34(2): 187-92, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12740577

ABSTRACT

DNA methylation inhibits gene expression in animal cells, probably by affecting chromatin structure. Biochemical studies suggest that this process may be mediated by methyl-specific binding proteins that recruit enzymatic machinery capable of locally altering histone modification. To test whether DNA methylation actually has a role in the assembly of chromatin during normal development, we used cell transfection and a transgene construct genetically programmed to be either methylated or unmethylated in all cell types of the mouse. Chromatin immunoprecipitation (ChIP) analysis shows that the presence of DNA methylation brings about the deacetylation of histone H4 and methylation of Lys9 of histone H3 (H3 Lys9) and prevents methylation of Lys4 of histone H3 (H3 Lys4), thus generating a structure identical to that of methylated sequences in the genome. These results indicate that the methylation pattern established in early embryogenesis is profoundly important in setting up the structural profile of the genome.


Subject(s)
Chromatin/genetics , Chromatin/metabolism , DNA Methylation , Acetylation , Animals , CpG Islands , Gene Expression Regulation, Developmental , Globins/genetics , Histones/chemistry , Histones/metabolism , Humans , Lysine/chemistry , Methylation , Mice , Mice, Transgenic , Models, Genetic , Transfection
11.
Nature ; 420(6912): 198-202, 2002 Nov 14.
Article in English | MEDLINE | ID: mdl-12432398

ABSTRACT

In animal cells, the process of DNA replication takes place in a programmed manner, with each gene region designated to replicate at a fixed time slot in S phase. Housekeeping genes undergo replication in the first half of S phase in all cell types, whereas the replication of many tissue specific genes is developmentally controlled, being late in most tissues but early in the tissue of expression. Here we employ nuclear DNA injection as an experimental system to test whether this phenomenon is due to differences in the ability to set up transcriptional competence during S phase. Our results show that, regardless of sequence, exogenous genes are a better template for transcription when injected into nuclei of cells in early as opposed to late S phase, and this expression state, once initiated, is preserved after cell division. DNA injected in late S phase is apparently repressed because it is packaged into chromatin containing deacetylated histones, and the same is true for late replicating chromosomal DNA. These findings suggest a mechanistic connection between replication timing and gene expression that might help to explain how epigenetic states can be maintained in vivo.


Subject(s)
Gene Expression Regulation , S Phase , Transcription, Genetic , Acetylation , Animals , Cell Division , Cell Line , Chromatin/chemistry , Chromatin/genetics , Chromatin/metabolism , Chromosomes/genetics , Chromosomes/metabolism , DNA/administration & dosage , DNA/chemistry , DNA/genetics , DNA/metabolism , DNA Replication , Gene Expression Regulation/drug effects , Gene Silencing/drug effects , Histone Deacetylase Inhibitors , Histone Deacetylases/metabolism , Histones/chemistry , Histones/metabolism , Hydroxamic Acids/pharmacology , Microinjections , Nucleic Acid Conformation , Organ Specificity , Rats , Templates, Genetic , Time Factors , Transcription, Genetic/drug effects
SELECTION OF CITATIONS
SEARCH DETAIL
...