MeCP2-chromatin interactions include the formation of chromatosome-like structures and are altered in mutations causing Rett syndrome.
hMeCP2 (human methylated DNA-binding protein 2), mutations of which cause most cases of Rett syndrome (RTT), is involved in the transmission of repressive epigenetic signals encoded by DNA methylation. The present work focuses on the modifications of chromatin architecture induced by MeCP2 and the effects of RTT-causing mutants. hMeCP2 binds to nucleosomes close to the linker DNA entry-exit site and protects approximately 11 bp of linker DNA from micrococcal nuclease. MeCP2 mutants differ in this property; the R106W mutant gives very little extra protection beyond the approximately 146-bp nucleosome core, whereas the large C-terminal truncation R294X reveals wild type behavior. Gel mobility assays show that linker DNA is essential for proper MeCP2 binding to nucleosomes, and electron microscopy visualization shows that the protein induces distinct conformational changes in the linker DNA. When bound to nucleosomes, MeCP2 is in close proximity to histone H3, which exits the nucleosome core close to the proposed MeCP2-binding site. These findings firmly establish nucleosomal linker DNA as a crucial binding partner of MeCP2 and show that different RTT-causing mutations of MeCP2 are correspondingly defective in different aspects of the interactions that alter chromatin architecture.
Department of Biology, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Tatiana
T
Nikitina
Rajarshi P
RP
Ghosh
Rachel A
RA
Horowitz-Scherer
Jeffrey C
JC
Hansen
Sergei A
SA
Grigoryev
Christopher L
CL
Woodcock
D001665:Binding Sites; D001667:Binding, Competitive; D002843:Chromatin / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry; D002875:Chromosomes / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D006657:Histones / Q000737:chemistry; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000737:chemistry / Q000502:physiology*; D008854:Microscopy, Electron; D008957:Models, Genetic; D009154:Mutation; D009707:Nucleosomes / Q000378:metabolism; D015518:Rett Syndrome / Q000235:genetics* / Q000378:metabolism
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D009707:Nucleosomes; D004247:DNA