Ballestar Esteban, Ropero Santiago, Alaminos Miguel, Armstrong Judith, Setien Fernando, Agrelo Ruben, Fraga Mario F, Herranz Michel, Avila Sonia, Pineda Mercedes, Monros Eugenia, Esteller Manel
Cancer Epigenetics Laboratory, Molecular Pathology Programme, Spanish National Cancer Centre, Melchor Fernández Almagro 3, 28029 Madrid, Spain.
Hum Genet. 2005 Jan;116(1-2):91-104. doi: 10.1007/s00439-004-1200-0. Epub 2004 Nov 11.
Rett syndrome (RTT), the second most common cause of mental retardation in females, has been associated with mutations in MeCP2, the archetypical member of the methyl-CpG binding domain (MBD) family of proteins. MeCP2 additionally possesses a transcriptional repression domain (TRD). We have compared the gene expression profiles of RTT- and normal female-derived lymphoblastoid cells by using cDNA microarrays. Clustering analysis allowed the classification of RTT patients according to the localization of the MeCP2 mutation (MBD or TRD) and those with clinically diagnosed RTT but without detectable MeCP2 mutations. Numerous genes were observed to be overexpressed in RTT patients compared with control samples, including excellent candidate genes for neurodevelopmental disease. Chromatin immunoprecipitation analysis confirmed that binding of MeCP2 to corresponding promoter CpG islands was lost in RTT-derived cells harboring a mutation in the region of the MECP2 gene encoding the MBD. Bisulfite genomic sequencing demonstrated that the majority of MeCP2 binding occurred in DNA sequences with methylation-associated silencing. Most importantly, the finding that these genes are also methylated and bound by MeCP2 in neuron-related cells suggests a role in this neurodevelopmental disease. Our results provide new data of the underlying mechanisms of RTT and unveil novel targets of MeCP2-mediated gene repression.
瑞特综合征(RTT)是女性智力发育迟缓的第二大常见病因,与甲基CpG结合结构域(MBD)蛋白家族的典型成员MeCP2突变有关。MeCP2还具有一个转录抑制结构域(TRD)。我们使用cDNA微阵列比较了源自RTT女性和正常女性的淋巴母细胞的基因表达谱。聚类分析能够根据MeCP2突变的定位(MBD或TRD)以及临床诊断为RTT但未检测到MeCP2突变的患者对RTT患者进行分类。与对照样本相比,观察到RTT患者中有许多基因过度表达,包括神经发育疾病的优秀候选基因。染色质免疫沉淀分析证实,在编码MBD的MECP2基因区域发生突变的RTT来源细胞中,MeCP2与相应启动子CpG岛的结合丧失。亚硫酸氢盐基因组测序表明,大多数MeCP2结合发生在与甲基化相关沉默的DNA序列中。最重要的是,这些基因在神经元相关细胞中也被甲基化并与MeCP2结合这一发现表明其在这种神经发育疾病中发挥作用。我们的结果提供了关于RTT潜在机制的新数据,并揭示了MeCP2介导的基因抑制的新靶点。