DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells

Spada, Fabio; Haemmer, Andrea; Kuch, David; Rothbauer, Ulrich; Schermelleh, Lothar; Kremmer, Elisabeth; Carell, Thomas; Längst, Gernot; Leonhardt, Heinrich

doi:10.1083/jcb.200610062

Cited by 171 publications

(149 citation statements)

References 30 publications

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“…For these experiments, we used DNMT1-knockout (DNMT1-KO) HCT116 colorectal cancer cells containing a deletion of the three exons encoding the DNMT1 regulatory domain. Although originally designed for a complete knockout, generation of a hypomorphic allele by alternative splicing allows expression of minute amounts of DNMT1 in these "DNMT1-null" HCT116 cells (19,62). Consistent with the earlier report (12), DNMT1-KO cells accumulated in G 2 phase in the absence of DNMT1, and treatment of these cells with DNA-damaging agents (gamma irradiation) further increases the percentage of cells in G 2 /M (Fig.…”

Section: Dnmt1 Is Acetylated In Vivo and In Vitrosupporting

confidence: 78%

SIRT1 Deacetylates the DNA Methyltransferase 1 (DNMT1) Protein and Alters Its Activities

Peng

Yuan

Ling

et al. 2011

Molecular and Cellular Biology

185

146

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DNA methylation and histone acetylation/deacetylation are distinct biochemical processes that control gene expression. While DNA methylation is a common epigenetic signal that inhibits gene transcription, histone deacetylation similarly represses transcription but can be both an epigenetic and nonepigenetic phenomenon. Here we report that the histone deacetylase SIRT1 regulates the activities of DNMT1, a key enzyme responsible for DNA methylation. In mass spectrometry analysis, 12 new acetylated lysine sites were identified in DNMT1. SIRT1 physically associates with DNMT1 and can deacetylate acetylated DNMT1 in vitro and in vivo. Interestingly, deacetylation of different lysines on DNMT1 has different effects on the functions of DNMT1. For example, deacetylation of Lys1349 and Lys1415 in the catalytic domain of DNMT1 enhances DNMT1's methyltransferase activity, while deacetylation of lysine residues in the GK linker decreases DNMT1's methyltransferase-independent transcriptional repression function. Furthermore, deacetylation of all identified acetylated lysine sites in DNMT1 abrogates its binding to SIRT1 and impairs its capability to regulate cell cycle G 2 /M transition. Finally, inhibition of SIRT1 strengthens the silencing effects of DNMT1 on the expression of tumor suppressor genes ER-␣ and CDH1 in MDA-MB-231 breast cancer cells. Together, these results suggest that SIRT1-mediated deacetylation of DNMT1 is crucial for DNMT1's multiple effects in gene silencing.

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Section: Dnmt1 Is Acetylated In Vivo and In Vitrosupporting

confidence: 78%

SIRT1 Deacetylates the DNA Methyltransferase 1 (DNMT1) Protein and Alters Its Activities

Peng

Yuan

Ling

et al. 2011

Molecular and Cellular Biology

185

146

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“…The level of processivity reaches up to 99% on hemimethylated DNA. Recent cell based DNA methylation studies of a PCNA binding mutant DNMT1s raises an interesting question about the absolute need of PCNA interaction with DNMT1 for DNA methylation propagation (40). Cells expressing a PCNA binding mutant of DNMT1 that does not load into the replication foci displayed a 2-fold lower DNA methylation efficiency (41).…”

Section: Discussionmentioning

confidence: 99%

CXXC Domain of Human DNMT1 Is Essential for Enzymatic Activity

et al. 2008

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DNA cytosine methylation is one of the major epigenetic gene silencing marks in the human genome facilitated by DNA methyltransferases. DNA cytosine-5 methyltransferase 1 (DNMT1) performs maintenance methylation in somatic cells. In cancer cells, DNMT1 is responsible for the aberrant hypermethylation of CpG islands and the silencing of tumor suppressor genes. Here we show that the catalytically active recombinant DNMT1, lacking 580 amino acids from the amino terminus, binds to unmethylated DNA with higher affinity than hemimethylated or methylated DNA. To further understand the binding domain of enzyme, we have used gel shift assay. We have demonstrated that the CXXC region (C is cysteine; X is any amino acid) of DNMT1 bound specifically to unmethylated CpG dinucleotides. Furthermore, mutation of the conserved cysteines abolished CXXC mediated DNA binding. In transfected COS-7 cells, CXXC deleted DNMT1 (DNMT1 ∆CXXC ) localized on replication foci. Both point mutant and DNMT1 ∆CXXC enzyme displayed significant reduction in catalytic activity, confirming that this domain is crucial for enzymatic activity. A permanent cell line with DNMT1 ∆CXXC displayed partial loss of genomic methylation on rDNA loci, despite the presence of endogenous wild-type enzyme. Thus, the CXXC domain encompassing the amino terminus region of DNMT1 cooperates with the catalytic domain for DNA methyltransferase activity.

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“…Moreover, the recruitment of DNMTs to specific gene targets suggests that our original picture of maintenance DNMT moving along with the replication fork [Gruenbaum et al, 1983;Leonhardt et al, 1992;Araujo et al, 1998] needs to be revisited. This challenge is further supported by recent data showing that a mutant DNMT1 defective in the replication fork targeting is still able to maintain most of the DNA methylation in the human genome [Spada et al, 2007]. Third, it is becoming clear that not only are DNMTs targeted to specific genes by sequence specific factors but they are also required to reside on these sequences to maintain their methylation state [Fuks et al, 2001;Di Croce et al, 2002;Brenner et al, 2005;Vire et al, 2006;Burgers et al, 2006].…”

Section: Dna Methylation Patterns and Their Maintenancementioning

confidence: 99%

The social environment and the epigenome

Szyf

McGowan

Meaney

2007

Environ and Mol Mutagen

454

222

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The genome is programmed by the epigenome. Two of the fundamental components of the epigenome are chromatin structure and covalent modification of the DNA molecule itself by methylation. DNA methylation patterns are sculpted during development and it has been a long held belief that they remain stable after birth in somatic tissues. Recent data suggest that DNA methylation is dynamic later in life in postmitotic cells such as neurons and thus potentially responsive to different environmental stimuli throughout life. We hypothesize a mechanism linking the social environment early in life and long-term epigenetic programming of behavior and responsiveness to stress and health status later in life. We will also discuss the prospect that the epigenetic equilibrium remains responsive throughout life and that therefore environmental triggers could play a role in generating interindividual differences in human behavior later in life. We speculate that exposures to different environmental toxins alters long-established epigenetic programs in the brain as well as other tissues leading to lateonset disease. Environ. Mol. Mutagen. 49:46-60, 2008. V V C 2007 Wiley-Liss, Inc.

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DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells

Cited by 171 publications

References 30 publications

SIRT1 Deacetylates the DNA Methyltransferase 1 (DNMT1) Protein and Alters Its Activities

SIRT1 Deacetylates the DNA Methyltransferase 1 (DNMT1) Protein and Alters Its Activities

CXXC Domain of Human DNMT1 Is Essential for Enzymatic Activity

The social environment and the epigenome

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