title: Unique physical properties and interactions of the domains of methylated DNA binding protein 2.
abstract: Methylated DNA binding protein 2 (MeCP2) is a methyl CpG binding protein whose key role is the recognition of epigenetic information encoded in DNA methylation patterns. Mutation or misregulation of MeCP2 function leads to Rett syndrome as well as a variety of other autism spectrum disorders. Here, we have analyzed in detail the properties of six individually expressed human MeCP2 domains spanning the entire protein with emphasis on their interactions with each other, with DNA, and with nucleosomal arrays. Each domain contributes uniquely to the structure and function of the full-length protein. MeCP2 is approximately 60% unstructured, with nine interspersed alpha-molecular recognition features (alpha-MoRFs), which are polypeptide segments predicted to acquire secondary structure upon forming complexes with binding partners. Large increases in secondary structure content are induced in some of the isolated MeCP2 domains and in the full-length protein by binding to DNA. Interactions between some MeCP2 domains in cis and trans seen in our assays likely contribute to the structure and function of the intact protein. We also show that MeCP2 has two functional halves. The N-terminal portion contains the methylated DNA binding domain (MBD) and two highly disordered flanking domains that modulate MBD-mediated DNA binding. One of these flanking domains is also capable of autonomous DNA binding. In contrast, the C-terminal portion of the protein that harbors at least two independent DNA binding domains and a chromatin-specific binding domain is largely responsible for mediating nucleosomal array compaction and oligomerization. These findings led to new mechanistic and biochemical insights regarding the conformational modulations of this intrinsically disordered protein, and its context-dependent in vivo roles.
authors.affiliation: Department of Biology, University of Massachusetts, Amherst, Massachusetts 01003, USA.
authors.forename: Rajarshi P
authors.initials: RP
authors.lastname: Ghosh
authors.forename: Tatiana
authors.initials: T
authors.lastname: Nikitina
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Horowitz-Scherer
authors.forename: Lila M
authors.initials: LM
authors.lastname: Gierasch
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Uversky
authors.forename: Kristopher
authors.initials: K
authors.lastname: Hite
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Hansen
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Woodcock
mesh_terms: D001665:Binding Sites; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology; D008958:Models, Molecular; D011485:Protein Binding / Q000502:physiology; D054730:Protein Interaction Domains and Motifs / Q000502:physiology*; D055550:Protein Stability; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary / Q000502:physiology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D002843:Chromatin; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D004247:DNA
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