Chromatin compaction by human MeCP2. Assembly of novel secondary chromatin structures in the absence of DNA methylation.
MeCP2 is a transcriptional repressor that contains an N-terminal methylated DNA-binding domain, a central transcription regulation domain, and a C-terminal domain of unknown function. Whereas current models of MeCP2 function evoke localized recruitment of histone deacetylases to specific methylated regions of the genome, it is unclear whether MeCP2 requires DNA methylation to bind to chromatin or whether MeCP2 binding influences chromatin structure in the absence of other proteins. To address these issues, we have characterized the complexes formed between MeCP2 and biochemically defined nucleosomal arrays. At molar ratios near 1 MeCP2/nucleosome, unmethylated nucleosomal arrays formed both extensively condensed ellipsoidal particles and oligomeric suprastructures. Furthermore, MeCP2-mediated chromatin compaction occurred in the absence of monovalent or divalent cations, in distinct contrast to all other known chromatin-condensing proteins. Analysis of specific missense and nonsense MeCP2 mutants indicated that the ability to condense chromatin resides in region(s) of the protein other than the methylated DNA-binding domain. These data demonstrate that MeCP2 assembles novel secondary chromatin structures independent of DNA modification and suggest that the ability of MeCP2 to silence chromatin may be related in part to its effects on large-scale chromatin organization.
Georgel
Philippe T
PT
Department of Biochemistry, University of Texas Health Science Center, San Antonio, Texas 78229-3900, USA.
Horowitz-Scherer
Rachel A
RA
Adkins
Nick
N
Woodcock
Christopher L
CL
Wade
Paul A
PA
Hansen
Jeffrey C
JC
D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D011994:Recombinant Proteins; D012097:Repressor Proteins