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PMID: 17101771 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Multiple modes of interaction between the methylated DNA binding protein MeCP2 and chromatin.

Molecular and cellular biology ·Vol. 27 ·No. 3 ·2007-02-00 ·Pages 864-77

Nikitina T, Shi X, Ghosh RP, Horowitz-Scherer RA, Hansen JC, Woodcock CL

Abstract

Mutations of the methylated DNA binding protein MeCP2, a multifunctional protein that is thought to transmit epigenetic information encoded as methylated CpG dinucleotides to the transcriptional machinery, give rise to the debilitating neurodevelopmental disease Rett syndrome (RTT). In this in vitro study, the methylation-dependent and -independent interactions of wild-type and mutant human MeCP2 with defined DNA and chromatin substrates were investigated. A combination of electrophoretic mobility shift assays and visualization by electron microscopy made it possible to understand the different conformational changes underlying the gel shifts. MeCP2 is shown to have, in addition to its well-established methylated DNA binding domain, a methylation-independent DNA binding site (or sites) in the first 294 residues, while the C-terminal portion of MeCP2 (residues 295 to 486) contains one or more essential chromatin interaction regions. All of the RTT-inducing mutants tested were quantitatively bound to chromatin under our conditions, but those that tend to be associated with the more severe RTT symptoms failed to induce the extensive compaction observed with wild-type MeCP2. Two modes of MeCP2-driven compaction were observed, one promoting nucleosome clustering and the other forming DNA-MeCP2-DNA complexes. MeCP2 binding to DNA and chromatin involves a number of different molecular interactions, some of which result in compaction and oligomerization. The multifunctional roles of MeCP2 may be reflected in these different interactions.

MeSH Terms
Chromatin/metabolism,ultrastructure DNA/chemistry,ultrastructure DNA Methylation Electrophoretic Mobility Shift Assay Humans Methyl-CpG-Binding Protein 2/chemistry,metabolism,ultrastructure Models, Biological Mutant Proteins/metabolism Nucleic Acid Conformation Nucleosomes/chemistry,ultrastructure Protein Binding Protein Conformation
Chemicals
Chromatin MECP2 protein, human Methyl-CpG-Binding Protein 2 Mutant Proteins Nucleosomes DNA
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nikitina Tatiana
Biology Department, University of Massachusetts, Amherst, MA 01003, USA.
Shi Xi
Ghosh Rajarshi P
Horowitz-Scherer Rachel A
Hansen Jeffrey C
Woodcock Christopher L
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.01593-06
Published
2007-02-00
Epub
2006-00-13
Pages
864-77
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1800686
Subset
IM
Grants
NIGMS NIH HHS · GM 070897 · United States
NIGMS NIH HHS · R01 GM066834 · United States
NIGMS NIH HHS · R01 GM070897 · United States
NIGMS NIH HHS · GM 45916 · United States
NIGMS NIH HHS · GM 66834 · United States
NIGMS NIH HHS · R01 GM045916 · United States
External Links
PubMed source DOI: 10.1128/MCB.01593-06 PMC Full Text View journal details
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