Source:http://linkedlifedata.com/resource/pubmed/id/18754681
Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
38
|
| pubmed:dateCreated |
2008-9-17
|
| pubmed:abstractText |
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 (DeltaCXXC)) localized on replication foci. Both point mutant and DNMT1 (DeltaCXXC) enzyme displayed significant reduction in catalytic activity, confirming that this domain is crucial for enzymatic activity. A permanent cell line with DNMT1 (DeltaCXXC) 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.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
1520-4995
|
| pubmed:author | |
| pubmed:issnType |
Electronic
|
| pubmed:day |
23
|
| pubmed:volume |
47
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
10000-9
|
| pubmed:meshHeading |
pubmed-meshheading:18754681-Amino Acid Motifs,
pubmed-meshheading:18754681-Amino Acid Sequence,
pubmed-meshheading:18754681-Animals,
pubmed-meshheading:18754681-Binding Sites,
pubmed-meshheading:18754681-COS Cells,
pubmed-meshheading:18754681-Catalytic Domain,
pubmed-meshheading:18754681-Cercopithecus aethiops,
pubmed-meshheading:18754681-Conserved Sequence,
pubmed-meshheading:18754681-Cysteine,
pubmed-meshheading:18754681-DNA (Cytosine-5-)-Methyltransferase,
pubmed-meshheading:18754681-DNA Methylation,
pubmed-meshheading:18754681-Enzyme Activation,
pubmed-meshheading:18754681-Humans,
pubmed-meshheading:18754681-Molecular Sequence Data,
pubmed-meshheading:18754681-Point Mutation,
pubmed-meshheading:18754681-Protein Folding,
pubmed-meshheading:18754681-Protein Structure, Tertiary,
pubmed-meshheading:18754681-Repetitive Sequences, Amino Acid,
pubmed-meshheading:18754681-Sequence Deletion,
pubmed-meshheading:18754681-Zinc Fingers
|
| pubmed:year |
2008
|
| pubmed:articleTitle |
CXXC domain of human DNMT1 is essential for enzymatic activity.
|
| pubmed:affiliation |
New England Biolabs, 240 County Road, Ipswich, Massachusetts 01938-2723, USA. pradhan@neb.com
|
| pubmed:publicationType |
Journal Article,
Comparative Study
|