Home LiteratureArticle Details
PMID: 8177735 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Dissection of the methyl-CpG binding domain from the chromosomal protein MeCP2.

Nucleic acids research ·Vol. 21 ·No. 21 ·1993-10-25 ·Pages 4886-92

Nan X, Meehan RR, Bird A

Abstract

MeCP2 is a chromosomal protein which binds to DNA that is methylated at CpG. In situ immunofluorescence in mouse cells has shown that the protein is most concentrated in pericentromeric heterochromatin, suggesting that MeCP2 may play a role in the formation of inert chromatin. Here we have isolated a minimal methyl-CpG binding domain (MBD) from MeCP2. MBD is 85 amino acids in length, and binds exclusively to DNA that contains one or more symmetrically methylated CpGs. MBD has negligable non-specific affinity for DNA, confirming that non-specific and methyl-CpG specific binding domains of MeCP2 are distinct. In vitro footprinting indicates that MBD binding can protect a 12 nucleotide region surrounding a methyl-CpG pair, with an approximate dissociation constant of 10(-9) M.

MeSH Terms
Base Sequence Binding Sites Chromosomal Proteins, Non-Histone DNA Primers DNA, Recombinant/metabolism DNA-Binding Proteins/chemistry,metabolism Dinucleoside Phosphates/metabolism Methyl-CpG-Binding Protein 2 Methylation Molecular Sequence Data Repressor Proteins Sequence Deletion
Chemicals
Chromosomal Proteins, Non-Histone DNA Primers DNA, Recombinant DNA-Binding Proteins Dinucleoside Phosphates Methyl-CpG-Binding Protein 2 Repressor Proteins cytidylyl-3'-5'-guanosine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nan X
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Meehan R R
Bird A
References (21)
21 references, click to expand
  1. Alternative processing of mRNAs encoding mammalian chromosomal high-mobility-group proteins HMG-I and HMG-Y.
    Mol Cell Biol. 1989 May;9(5):2114-23 PMID: 2701943
  2. A dimer of AraC protein contacts three adjacent major groove regions of the araI DNA site.
    Proc Natl Acad Sci U S A. 1985 May;82(10):3129-33 PMID: 3858809
  3. Specific protection of methylated CpGs in mammalian nuclei.
    Cell. 1989 Aug 11;58(3):509-17 PMID: 2474378
  4. 'SPKK' motifs prefer to bind to DNA at A/T-rich sites.
    EMBO J. 1989 Dec 20;8(13):4189-95 PMID: 2556263
  5. DNA methylation and chromatin structure: a view from below.
    Trends Biochem Sci. 1990 Mar;15(3):103-7 PMID: 2139257
  6. The A.T-DNA-binding domain of mammalian high mobility group I chromosomal proteins. A novel peptide motif for recognizing DNA structure.
    J Biol Chem. 1990 May 25;265(15):8573-82 PMID: 1692833
  7. Use of T7 RNA polymerase to direct expression of cloned genes.
    Methods Enzymol. 1990;185:60-89 PMID: 2199796
  8. In vivo footprint and methylation analysis by PCR-aided genomic sequencing: comparison of active and inactive X chromosomal DNA at the CpG island and promoter of human PGK-1.
    Genes Dev. 1990 Aug;4(8):1277-87 PMID: 2227409
  9. DNA methylation inhibits transcription indirectly via a methyl-CpG binding protein.
    Cell. 1991 Mar 22;64(6):1123-34 PMID: 2004419
  10. Chromatin differences between active and inactive X chromosomes revealed by genomic footprinting of permeabilized cells using DNase I and ligation-mediated PCR.
    Genes Dev. 1991 Jun;5(6):1102-13 PMID: 2044957
  11. DNA methylation and chromatin structure.
    FEBS Lett. 1991 Jul 22;285(2):155-9 PMID: 1855583
  12. Diversity and specificity in transcriptional regulation: the benefits of heterotypic dimerization.
    Trends Biochem Sci. 1991 Nov;16(11):417-22 PMID: 1776171
  13. Purification, sequence, and cellular localization of a novel chromosomal protein that binds to methylated DNA.
    Cell. 1992 Jun 12;69(6):905-14 PMID: 1606614
  14. Identification of an amino acid-base contact in the GCN4-DNA complex by bromouracil-mediated photocrosslinking.
    Nature. 1992 Oct 15;359(6396):650-2 PMID: 1406998
  15. Characterization of MeCP2, a vertebrate DNA binding protein with affinity for methylated DNA.
    Nucleic Acids Res. 1992 Oct 11;20(19):5085-92 PMID: 1408825
  16. The GCN4 basic region leucine zipper binds DNA as a dimer of uninterrupted alpha helices: crystal structure of the protein-DNA complex.
    Cell. 1992 Dec 24;71(7):1223-37 PMID: 1473154
  17. Transcriptional repression by methylation of CpG.
    J Cell Sci Suppl. 1992;16:9-14 PMID: 1297654
  18. Mr 26,000 antigen of Schistosoma japonicum recognized by resistant WEHI 129/J mice is a parasite glutathione S-transferase.
    Proc Natl Acad Sci U S A. 1986 Nov;83(22):8703-7 PMID: 3095841
  19. 5-Methylcytosine localised in mammalian constitutive heterochromatin.
    Nature. 1974 Oct 18;251(5476):636-7 PMID: 4609195
  20. Nucleosomes are positioned on mouse satellite DNA in multiple highly specific frames that are correlated with a diverged subrepeat of nine base-pairs.
    J Mol Biol. 1984 Jun 15;176(1):105-29 PMID: 6330367
  21. Identification of a mammalian protein that binds specifically to DNA containing methylated CpGs.
    Cell. 1989 Aug 11;58(3):499-507 PMID: 2758464
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/21.21.4886
Published
1993-10-25
Pages
4886-92
Language
English
Region
England
NLM ID
0411011
PMCID
PMC311401
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/21.21.4886 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2