Division of Pediatrics, Dokkyo University Koshigaya Hospital, 2-1-50 Minami Koshigaya, Koshigaya, Saitama 343-8555, Japan.
Hum Genet. 2012 Jan;131(1):121-30. doi: 10.1007/s00439-011-1051-4. Epub 2011 Jul 7.
X-chromosome inactivation (XCI) is an essential mechanism in females that compensates for the genome imbalance between females and males. It is known that XCI can spread into an autosome of patients with X;autosome translocations. The subject was a 5-year-old boy with Prader-Willi syndrome (PWS)-like features including hypotonia, hypo-genitalism, hypo-pigmentation, and developmental delay. G-banding, fluorescent in situ hybridization, BrdU-incorporated replication, human androgen receptor gene locus assay, SNP microarrays, ChIP-on-chip assay, bisulfite sequencing, and real-time RT-PCR were performed. Cytogenetic analyses revealed that the karyotype was 46,XY,der(X)t(X;15)(p21.1;q11.2),-15. In the derivative chromosome, the X and half of the chromosome 15 segments showed late replication. The X segment was maternal, and the chromosome 15 region was paternal, indicating its post-zygotic origin. The two chromosome 15s had a biparental origin. The DNA methylation level was relatively high in the region proximal from the breakpoint, and the level decreased toward the middle of the chromosome 15 region; however, scattered areas of hypermethylation were found in the distal region. The promoter regions of the imprinted SNRPN and the non-imprinted OCA2 genes were completely and half methylated, respectively. However, no methylation was found in the adjacent imprinted gene UBE3A, which contained a lower density of LINE1 repeats. Our findings suggest that XCI spread into the paternal chromosome 15 led to the aberrant hypermethylation of SNRPN and OCA2 and their decreased expression, which contributes to the PWS-like features and hypo-pigmentation of the patient. To our knowledge, this is the first chromosome-wide methylation study in which the DNA methylation level is demonstrated in an autosome subject to XCI.
X 染色体失活(XCI)是女性中一种重要的机制,可补偿女性和男性之间的基因组不平衡。已知 XCI 可以扩展到具有 X;常染色体易位的患者的常染色体中。研究对象是一名 5 岁男孩,具有 Prader-Willi 综合征(PWS)样特征,包括低张力、性腺发育不全、色素减退和发育迟缓。进行了 G 带、荧光原位杂交、BrdU 掺入复制、人雄激素受体基因座测定、SNP 微阵列、ChIP-on-chip 测定、亚硫酸氢盐测序和实时 RT-PCR。细胞遗传学分析显示核型为 46,XY,der(X)t(X;15)(p21.1;q11.2),-15。在衍生染色体中,X 染色体和 15 号染色体的一半片段显示晚期复制。X 染色体片段为母系,15 号染色体区域为父系,表明其为合子后起源。两条 15 号染色体具有双亲的起源。靠近断裂点的近端区域的 DNA 甲基化水平相对较高,向 15 号染色体区域的中间降低;然而,在远端发现了散在的高甲基化区域。印迹 SNRPN 和非印迹 OCA2 基因的启动子区域分别完全和半甲基化,而相邻的印迹基因 UBE3A 则未发现甲基化,其含有较低密度的 LINE1 重复序列。我们的研究结果表明,XCI 扩展到父系 15 号染色体导致 SNRPN 和 OCA2 的异常高甲基化及其表达降低,这导致患者出现 PWS 样特征和色素减退。据我们所知,这是首次在受 XCI 影响的常染色体上进行全染色体甲基化研究,证明了 DNA 甲基化水平。