Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA177404 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR586845.fastq.gz | Fastqsanger.gz |
Items per page: 5 1 - 1 of 1 |
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Project description:The TET3 CXXC domain has unique DNA binding properties. It binds to DNA in a cytosine-dependent manner that prefers binding to CpG dinucleotides but is not restricted by the CpG-content, distinct from other well-characterized CXXC domains. To map the TET3 CXXC domain binding sites across the human genome, we purified the GST-tagged TET3 CXXC domain protein and performed the GST pull-down assay using the genomic DNA purified from HEK293T cells. The enriched DNA fragments were then sequenced and aligned to human genome(hg19). We used the GST pull-down assay followed by DNA deep sequencing to map the DNA bound by the TET3 CXXC domain in vitro.
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Project description:The TET3 CXXC domain has unique DNA binding properties. It binds to DNA in a cytosine-dependent manner that prefers binding to CpG dinucleotides but is not restricted by the CpG-content, distinct from other well-characterized CXXC domains. To map the TET3 CXXC domain binding sites across the human genome, we purified the GST-tagged TET3 CXXC domain protein and performed the GST pull-down assay using the genomic DNA purified from HEK293T cells. The enriched DNA fragments were then sequenced and aligned to human genome(hg19).
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Project description:We report that the 5-methylcytosine oxidase Tet3 exists as three isoforms and characterized the full-length isoform containing an N-terminal CXXC domain (Tet3FL). Presence of the CXXC domain reduces Tet3’s enzymatic activity. This CXXC domain binds not only to unmethylated CpGs but its highest affinity is towards 5-carboxylcytosine (5caC). We determined the structure of the CXXC domain - 5caC-DNA complex and showed that disruption of the CXXC-5caC interaction leads to enhanced Tet3 activity. Mapping of genomic binding sites of Tet3FL in neuronal cells shows that Tet3FL is targeted to transcription start sites of genes involved in lysosome function, mRNA processing and key genes of the base excision repair pathway. Our data suggest that the CXXC domain of Tet3FL restricts the protein to a set of unmethylated or 5caC-containing CpG islands and that Tet3FL functions both as a reader of 5caC and as a regulator of 5caC removal by base excision repair.
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Project description:Idax/Cxxc4 is the CXXC domain protein that preferentially binds unmethylated CpG sites in vitro. By performing genome-wide mapping of Idax/Cxxc4, we report here that Idax is preferentially enriched in CpG-rich regions in genome including the promoters containing CpG islands.
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Project description:Idax/Cxxc4 is the CXXC domain protein that preferentially binds unmethylated CpG sites in vitro. By performing genome-wide mapping of Idax/Cxxc4, we report here that Idax is preferentially enriched in CpG-rich regions in genome including the promoters containing CpG islands. ChIP-seq binding site detection of Myc-Idax and Myc-IdaxDBM over cells expressing an empty vector in stable HEK293T cell lines; two replicates per condition (2x ChIP Myc-Idax, 2x ChIP Myc-IdaxDBM, 2xChIP empty vector)
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