title: Heterogeneity in residual function of MeCP2 carrying missense mutations in the methyl CpG binding domain.
abstract: Rett syndrome is a neurodevelopmental disorder with severe mental retardation caused by mutations in the MECP2 gene. Mutations in the MECP2 gene are also associated with other genetic disorders, including X linked mental retardation in males. Missense mutations identified so far are present primarily in the methyl CpG binding domain (MBD) of MECP2. Here, the functional significance of 28 MBD missense mutations identified in patients were analysed by transient expression of the mutant proteins in cultured cells. The effects of mutations were evaluated by analysis of the affinity of MeCP2 to pericentromeric heterochromatin in mouse L929 cells and on transcriptional repressive activity of MeCP2 in Drosophila SL2 cells. These analyses showed that approximately one-third (9/28) of MBD missense mutations showed strong impairment of MeCP2 function. The mutation of the R111 residue, which directly interacts with the methyl group of methyl cytosine, completely abolished MeCP2 function and mutations affecting beta-sheets and a hairpin loop have substantial functional consequences. In contrast, mutations that showed marginal or mild impairment of the function fell in unstructured regions with no DNA interaction. Since each of these mutations is known to be pathogenic, the mutations may indicate residues that are important for specific functions of MeCP2 in neurones.
authors.affiliation: Hokkaido Institute of Public Health, Sapporo 060-0819, Japan. kudos@iph.pref.hokkaido.jp
authors.forename: S
authors.initials: S
authors.lastname: Kudo
authors.forename: Y
authors.initials: Y
authors.lastname: Nomura
authors.forename: M
authors.initials: M
authors.lastname: Segawa
authors.forename: N
authors.initials: N
authors.lastname: Fujita
authors.forename: M
authors.initials: M
authors.lastname: Nakao
authors.forename: C
authors.initials: C
authors.lastname: Schanen
authors.forename: M
authors.initials: M
authors.lastname: Tamura
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive; D002460:Cell Line; D002868:Chromosomal Proteins, Non-Histone; D018899:CpG Islands / Q000235:genetics*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D018740:Genetic Heterogeneity; D049452:Green Fluorescent Proteins; D006570:Heterochromatin / Q000378:metabolism; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051783:Methyl-CpG-Binding Protein 2; D051379:Mice; D008856:Microscopy, Fluorescence; D020125:Mutation, Missense; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins; D015518:Rett Syndrome / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006570:Heterochromatin; D008164:Luminescent Proteins; C497392:MECP2 protein, human; C497393:Mecp2 protein, mouse; D051783:Methyl-CpG-Binding Protein 2; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D049452:Green Fluorescent Proteins