Characterization of MeCP2, a vertebrate DNA binding protein with affinity for methylated DNA.
Methylated DNA in vertebrates is associated with transcriptional repression and inactive chromatin. Two activities have been identified, MeCP1 and MeCP2, which bind specifically to DNA containing methyl-CpG pairs. In this report we characterize MeCP2. We show that it is more abundant than MeCP1, is more tightly bound in the nucleus, and is distinguishable chromatographically. The two proteins share widespread expression in somatic mammalian cells, and barely detectable expression in early embryonic cells. DNAs containing thymidine which has a methyl group at position 5 are not ligands for the MeCPs. The possible role of MeCP2 in methylation-associated gene inactivation was tested in in vitro transcription extracts. Purified MeCP2 inhibited transcription from both methylated and nonmethylated DNA templates in vitro, probably due to the presence of nonspecific DNA binding domains within the protein. We hypothesise that MeCP2 normally binds methylated DNA in the context of chromatin, contributing to the long-term repression and nuclease-resistance of methyl-CpGs.
Meehan
R R
RR
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Lewis
J D
JD
Bird
A P
AP
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D002846:Chromatography, Affinity; D002868:Chromosomal Proteins, Non-Histone; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006655:Histone Deacetylases; D015151:Immunoblotting; D007202:Indicators and Reagents; D051783:Methyl-CpG-Binding Protein 2; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D012097:Repressor Proteins; D013379:Substrate Specificity; D014158:Transcription, Genetic
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D007202:Indicators and Reagents; C489690:MeCP1 histone deacetylase complex, mouse; C077490:MeCP1 histone deacetylase complex, rat; C497393:Mecp2 protein, mouse; C497394:Mecp2 protein, rat; D051783:Methyl-CpG-Binding Protein 2; D012097:Repressor Proteins; D004247:DNA; D006655:Histone Deacetylases