Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 2 de 2
Filter
Add more filters










Database
Language
Publication year range
1.
Proc Natl Acad Sci U S A ; 108(14): 5718-23, 2011 Apr 05.
Article in English | MEDLINE | ID: mdl-21427230

ABSTRACT

Methylation on lysine 9 of histone H3 (H3K9me) and DNA methylation play important roles in the transcriptional silencing of specific genes and repetitive elements. Both marks are detected on class I and II endogenous retroviruses (ERVs) in murine embryonic stem cells (mESCs). Recently, we reported that the H3K9-specific lysine methyltransferase (KMTase) Eset/Setdb1/KMT1E is required for H3K9me3 and the maintenance of silencing of ERVs in mESCs. In contrast, G9a/Ehmt2/KMT1C is dispensable, despite the fact that this KMTase is required for H3K9 dimethylation (H3K9me2) and efficient DNA methylation of these retroelements. Transcription of the exogenous retrovirus (XRV) Moloney murine leukemia virus is rapidly extinguished after integration in mESCs, concomitant with de novo DNA methylation. However, the role that H3K9 KMTases play in this process has not been addressed. Here, we demonstrate that G9a, but not Suv39h1 or Suv39h2, is required for silencing of newly integrated Moloney murine leukemia virus-based vectors in mESCs. The silencing defect in G9a(-/-) cells is accompanied by a reduction of H3K9me2 at the proviral LTR, indicating that XRVs are direct targets of G9a. Furthermore, de novo DNA methylation of newly integrated proviruses is impaired in the G9a(-/-) line, phenocopying proviral DNA methylation and silencing defects observed in Dnmt3a-deficient mESCs. Once established, however, maintenance of silencing of XRVs, like ERVs, is dependent exclusively on the KMTase Eset. Taken together, these observations reveal that in mESCs, the H3K9 KMTase G9a is required for the establishment, but not for the maintenance, of silencing of newly integrated proviruses.


Subject(s)
DNA Methylation/genetics , Embryonic Stem Cells/virology , Histone-Lysine N-Methyltransferase/metabolism , Moloney murine leukemia virus/genetics , Animals , Blotting, Western , Chromatin Immunoprecipitation , DNA (Cytosine-5-)-Methyltransferases/genetics , Endogenous Retroviruses/genetics , Flow Cytometry , Gene Silencing , Genetic Vectors/genetics , Histone-Lysine N-Methyltransferase/genetics , Mice , Mice, Knockout , Proviruses/genetics , Reverse Transcriptase Polymerase Chain Reaction
2.
EMBO J ; 27(20): 2691-701, 2008 Oct 22.
Article in English | MEDLINE | ID: mdl-18818693

ABSTRACT

Histone H3K9 methylation is required for DNA methylation and silencing of repetitive elements in plants and filamentous fungi. In mammalian cells however, deletion of the H3K9 histone methyltransferases (HMTases) Suv39h1 and Suv39h2 does not affect DNA methylation of the endogenous retrovirus murine leukaemia virus, indicating that H3K9 methylation is dispensable for DNA methylation of retrotransposons, or that a different HMTase is involved. We demonstrate that embryonic stem (ES) cells lacking the H3K9 HMTase G9a show a significant reduction in DNA methylation of retrotransposons, major satellite repeats and densely methylated CpG-rich promoters. Surprisingly, demethylated retrotransposons remain transcriptionally silent in G9a(-/-) cells, and show only a modest decrease in H3K9me2 and no decrease in H3K9me3 or HP1alpha binding, indicating that H3K9 methylation per se is not the relevant trigger for DNA methylation. Indeed, introduction of catalytically inactive G9a transgenes partially 'rescues' the DNA methylation defect observed in G9a(-/-) cells. Taken together, these observations reveal that H3K9me3 and HP1alpha recruitment to retrotransposons occurs independent of DNA methylation in ES cells and that G9a promotes DNA methylation independent of its HMTase activity.


Subject(s)
DNA Methylation , Embryonic Stem Cells/cytology , Gene Expression Regulation, Enzymologic , Histone-Lysine N-Methyltransferase/metabolism , Methyltransferases/metabolism , Animals , Catalysis , Chromobox Protein Homolog 5 , Chromosomal Proteins, Non-Histone/metabolism , CpG Islands , Histones/chemistry , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Transgenic , Models, Genetic
SELECTION OF CITATIONS
SEARCH DETAIL
...