Unknown,Transcriptomics,Genomics,Proteomics

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Effect of TET1 overexpression on DNA methylation levels of HEK293 cells

ABSTRACT: We compared TET1 overexpressing cells to uninduced cells with endogenous TET1 transcript levels to determine DNA methylation changes. TET1 overexpression

ORGANISM(S): Homo sapiens

SUBMITTER: Christian Grosser

PROVIDER: E-GEOD-67349 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Altering TET dioxygenase levels within physiological range affects DNA methylation dynamics of HEK293 cells.

Grosser Christian C Wagner Nicholas N Grothaus Katrin K Horsthemke Bernhard B

Epigenetics 20150717 9

The TET family of dioxygenases (TET1/2/3) can convert 5-methylcytosine (5 mC) into 5-hydroxymethylcytosine (5 hmC) and has been shown to be involved in active and passive DNA demethylation. Here, we demonstrate that altering TET dioxygenase levels within physiological range can affect DNA methylation dynamics of HEK293 cells. Overexpression of TET1 increased global 5 hmC levels and was accompanied by mild DNA demethylation of promoters, gene bodies and CpG islands. Conversely, the simultaneous k ...[more]

PMID: 26186463

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Project description:Epigenetic modification of the mammalian genome by DNA methylation (5-methylcytosine) has a profound impact on chromatin structure, gene expression and maintenance of cellular identity. Recent demonstration that members of the Ten-eleven translocation (Tet) family proteins can convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) raised the possibility that Tet proteins are capable of establishing a distinct epigenetic state. We have recently demonstrated that Tet1 is specifically expressed in murine embryonic stem (ES) cells and is required for ES cell self-renewal and maintenance. Using chromatin immunoprecipitation coupled with high-throughput DNA sequencing (ChIP-seq), here we show that Tet1 is preferentially bound to CpG-rich sequences at promoters of both transcriptionally active and Polycomb-repressed genes. Despite a general increase in levels of DNA methylation at Tet1 binding-sites, Tet1 depletion does not lead to down-regulation of all the Tet1 targets. Interestingly, while Tet1-mediated promoter hypomethylation is required for maintaining the expression of a group of transcriptionally active genes, it is also required for repression of Polycomb-targeted developmental regulators. Tet1 contributes to silencing of this group of genes by facilitating recruitment of PRC2 to CpG-rich gene promoters. Thus, our study not only establishes a role for Tet1 in modulating DNA methylation levels at CpG-rich promoters, but also reveals a dual function of Tet1 in promoting transcription of pluripotency factors as well as participating in the repression of Polycomb-targeted developmental regulators. Mouse ES cells infected with control knockdown (KD) or Tet1 KD lentiviruses were FACS-sorted for RNA extraction and hybridization on Affymetrix microarrays. We also investigated the effect of Nanog overexpression (OE) in Tet1 KD mouse ES cells on dys-regulated Tet1 targets. We have collected four biologically independent replicates for each treatment.

Dataset Information

Effect of TET1 overexpression on DNA methylation levels of HEK293 cells

Publications

Altering TET dioxygenase levels within physiological range affects DNA methylation dynamics of HEK293 cells.

Similar Datasets

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