Genomics

Dataset Information

348

TET1 is a maintenance DNA demethylase that prevents methylation spreading in adult cells [DREAM]


ABSTRACT: We report that full length TET1 (TET1-FL) overexpression fails to induce global DNA demethylation in HEK293T cells. The preferential binding of TET1-FL to hypomethylated CpG islands (CGIs) through its CXXC domain leads to its inhibited 5-hydroxymethylcytosine (5hmC) production as methylation level increases. TET1-FL-induced 5hmC accumulates at CGI edges, while TET1 knockdown induces methylation spreading from methylated edges into hypomethylated CGIs. However, TET1 can regulate gene transcription independent of its dioxygenase catalytic function. Thus, our results identify TET1 as a maintenance DNA demethylase that does not purposely decrease methylation levels, but specifically maintains the DNA hypomethylation state of CGIs in adult cells. Digital restriction enzyme analysis of methylation (DREAM) was performed to determine the DNA methylation profiles of HEK293T cells overexpressing mTET1-CD, TET1-CD, mTET1-FL, or TET1-FL. In this approach, genomic DNA is sequentially cut at CCCGGG sites with the methylation sensitive enzyme SmaI (blunt ends) and its methylation-tolerant neoschizomer XmaI (5’CCGG overhangs), creating different end sequences that represent methylation status of the CCCGGG sites. These end sequences are analyzed by next generation sequencing, and thereafter the methylation status at individual CCCGGG sites across the genome can be determined.

ORGANISM(S): Homo sapiens  

SUBMITTER: CHUNLEI JIN   Michelle Barton  Shoudan Liang  Jaroslav Jelinek  Jean-Pierre J Issa  Chunlei Jin  Yue Lu  Marocs R Estecio 

PROVIDER: E-GEOD-44038 | ArrayExpress | 2014-06-04

SECONDARY ACCESSION(S): GSE44038SRP018556PRJNA188410

REPOSITORIES: GEO, ArrayExpress, ENA

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Publications

TET1 is a maintenance DNA demethylase that prevents methylation spreading in differentiated cells.

Jin Chunlei C   Lu Yue Y   Jelinek Jaroslav J   Liang Shoudan S   Estecio Marcos R H MR   Barton Michelle Craig MC   Issa Jean-Pierre J JP  

Nucleic acids research 20140529 11


TET1 is a 5-methylcytosine dioxygenase and its DNA demethylating activity has been implicated in pluripotency and reprogramming. However, the precise role of TET1 in DNA methylation regulation outside of developmental reprogramming is still unclear. Here, we show that overexpression of the TET1 catalytic domain but not full length TET1 (TET1-FL) induces massive global DNA demethylation in differentiated cells. Genome-wide mapping reveals that 5-hydroxymethylcytosine production by TET1-FL is inhi  ...[more]

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