title: Biophysical analysis and small-angle X-ray scattering-derived structures of MeCP2-nucleosome complexes.
abstract: MeCP2 is a highly abundant chromatin architectural protein with key roles in post-natal brain development in humans. Mutations in MeCP2 are associated with Rett syndrome, the main cause of mental retardation in girls. Structural information on the intrinsically disordered MeCP2 protein is restricted to the methyl-CpG binding domain; however, at least four regions capable of DNA and chromatin binding are distributed over its entire length. Here we use small angle X-ray scattering (SAXS) and other solution-state approaches to investigate the interaction of MeCP2 and a truncated, disease-causing version of MeCP2 with nucleosomes. We demonstrate that MeCP2 forms defined complexes with nucleosomes, in which all four histones are present. MeCP2 retains an extended conformation when binding nucleosomes without extra-nucleosomal DNA. In contrast, nucleosomes with extra-nucleosomal DNA engage additional DNA binding sites in MeCP2, resulting in a rather compact higher-order complex. We present ab initio envelope reconstructions of nucleosomes and their complexes with MeCP2 from SAXS data. SAXS studies also revealed unexpected sequence-dependent conformational variability in the nucleosomes themselves.
authors.affiliation: Department of Biochemistry and Molecular Biology and Howard Hughes Medical Institute, Colorado State University, Fort Collins, CO 80523-1870, USA.
authors.forename: Chenghua
authors.initials: C
authors.lastname: Yang
authors.forename: Mark J
authors.initials: MJ
authors.lastname: van der Woerd
authors.forename: Uma M
authors.initials: UM
authors.lastname: Muthurajan
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Hansen
authors.forename: Karolin
authors.initials: K
authors.lastname: Luger
mesh_terms: D001665:Binding Sites; D004247:DNA / Q000737:chemistry; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry* / Q000378:metabolism; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
chemical_list: C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D009707:Nucleosomes; D004247:DNA