Nucleic Acids Research, 1993, Vol. 21, No. 21 4886-4892
© 1993
MOLECULAR BIOLOGY |
Dissection of the methyl-CpG binding domain from the chromosomal protein MeCP2
Institute of Cell and Molecular Biology, University of Edinburgh Kings Buildings, Mayfield Road, Edinburgh EH9 3JR, UK
*To whom correspondence should be addressed
Received August 5, 1993. Revised September 9, 1993. Accepted September 9, 1993.
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 10–9M.
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J. P. Cheadle, H. Gill, N. Fleming, J. Maynard, A. Kerr, H. Leonard, M. Krawczak, D. N. Cooper, S. Lynch, N. Thomas, et al. Long-read sequence analysis of the MECP2 gene in Rett syndrome patients: correlation of disease severity with mutation type and location Hum. Mol. Genet., April 12, 2000; 9(7): 1119 - 1129. [Abstract] [Full Text] [PDF]
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F. Xiang, S. Buervenich, P. Nicolao, M. E S Bailey, Z. Zhang, and M. Anvret Mutation screening in Rett syndrome patients J. Med. Genet., April 1, 2000; 37(4): 250 - 255. [Abstract] [Full Text]
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H.-H. Ng, P. Jeppesen, and A. Bird Active Repression of Methylated Genes by the Chromosomal Protein MBD1 Mol. Cell. Biol., February 15, 2000; 20(4): 1394 - 1406. [Abstract] [Full Text]
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I. P. Pogribny, M. Pogribna, J. K. Christman, and S. J. James Single-Site Methylation within the p53 Promoter Region Reduces Gene Expression in a Reporter Gene Construct: Possible in Vivo Relevance during Tumorigenesis Cancer Res., February 1, 2000; 60(3): 588 - 594. [Abstract] [Full Text]
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A Akhmanova, T Verkerk, A Langeveld, F Grosveld, and N Galjart Characterisation of transcriptionally active and inactive chromatin domains in neurons J. Cell Sci., January 12, 2000; 113(24): 4463 - 4474. [Abstract] [PDF]
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N. Fujita, S.-i. Takebayashi, K. Okumura, S. Kudo, T. Chiba, H. Saya, and M. Nakao Methylation-Mediated Transcriptional Silencing in Euchromatin by Methyl-CpG Binding Protein MBD1 Isoforms Mol. Cell. Biol., September 1, 1999; 19(9): 6415 - 6426. [Abstract] [Full Text] [PDF]
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Y. Zhang, H.-H. Ng, H. Erdjument-Bromage, P. Tempst, A. Bird, and D. Reinberg Analysis of the NuRD subunits reveals a histone deacetylase core complex and a connection with DNA methylation Genes & Dev., August 1, 1999; 13(15): 1924 - 1935. [Abstract] [Full Text]
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M. Shiraishi, Y. H. Chuu, and T. Sekiya Isolation of DNA fragments associated with methylated CpG islands in human adenocarcinomas of the lung using a methylated DNA binding column and denaturing gradient gel electrophoresis PNAS, March 16, 1999; 96(6): 2913 - 2918. [Abstract] [Full Text] [PDF]
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B. Hendrich and A. Bird Identification and Characterization of a Family of Mammalian Methyl-CpG Binding Proteins Mol. Cell. Biol., November 1, 1998; 18(11): 6538 - 6547. [Abstract] [Full Text]
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S. Kudo Methyl-CpG-Binding Protein MeCP2 Represses Sp1-Activated Transcription of the Human Leukosialin Gene When the Promoter Is Methylated Mol. Cell. Biol., September 1, 1998; 18(9): 5492 - 5499. [Abstract] [Full Text]
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P.A. WADE, P.L. JONES, D. VERMAAK, G.J.C. VEENSTRA, A. IMHOF, T. SERA, C. TSE, H. GE, Y.-B. SHI, J.C. HANSEN, et al. Histone Deacetylase Directs the Dominant Silencing of Transcription in Chromatin: Association with MeCP2 and the Mi-2 Chromodomain SWI/SNF ATPase Cold Spring Harb Symp Quant Biol, January 1, 1998; 63(0): 435 - 446. [Abstract] [PDF]
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F. J. Campoy, R. R. Meehan, S. McKay, J. Nixon, and A. Bird Binding of Histone H1 to DNA Is Indifferent to Methylation at CpG Sequences J. Biol. Chem., November 3, 1995; 270(44): 26473 - 26481. [Abstract] [Full Text] [PDF]
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