Gu Y H, Kodama H, Murata Y, Mochizuki D, Yanagawa Y, Ushijima H, Shiba T, Lee C C
Department of Pediatrics, Teikyo University School of Medicine, Tokyo, Japan.
Am J Med Genet. 2001 Mar 15;99(3):217-22. doi: 10.1002/1096-8628(2001)9999:9999<::aid-ajmg1167>3.0.co;2-r.
Genomic DNA of 17 unrelated Japanese males with Menkes disease and 2 Japanese males with occipital horn syndrome were studied for mutations in the ATP7A gene. Using SSCP analysis and direct sequencing of the exons and the 5'-upstream region of the gene amplified by PCR, we identified 16 mutations in 16 of 17 males with Menkes disease, including 4 deletions, 2 insertions, 6 nonsense mutations, 2 missense mutations, and 2 splice-site mutations. All these mutations were those that affect the function of the gene. Of the two males with occipital horn syndrome, one had a splice-site mutation in intron 6 that led to normal-size and smaller-size transcripts. The amount of the normal-size transcripts in his cultured skin fibroblasts was 19% of the normal level. His serum copper and ceruloplasmin levels were normal, whereas his cultured skin fibroblasts contained increased levels of copper. These findings indicate that his mild clinical manifestations were due to the presence of normal-size and presumably functional transcripts of the gene. DNA sequencing analysis of the exons and 5'-upstream region of the ATP7A gene in 20 normal individuals and the 19 affected males identified 25 polymorphisms.
对17名患门克斯病的不相关日本男性和2名患枕角综合征的日本男性的基因组DNA进行了研究，以检测ATP7A基因中的突变。通过单链构象多态性（SSCP）分析以及对经聚合酶链反应（PCR）扩增的该基因外显子和5'-上游区域进行直接测序，我们在17名患门克斯病的男性中的16名中鉴定出16种突变，包括4种缺失、2种插入、6种无义突变、2种错义突变和2种剪接位点突变。所有这些突变均为影响该基因功能的突变。在两名患枕角综合征的男性中，其中一名在第6内含子中有一个剪接位点突变，导致产生正常大小和较小大小的转录本。在他培养的皮肤成纤维细胞中，正常大小转录本的量为正常水平的19%。他的血清铜和铜蓝蛋白水平正常，而他培养的皮肤成纤维细胞中铜含量增加。这些发现表明，他的轻度临床表现是由于该基因存在正常大小且可能具有功能的转录本。对20名正常个体和19名患病男性的ATP7A基因外显子和5'-上游区域进行DNA测序分析，鉴定出25种多态性。
