Qi M, Byers P H
Department of Pathology, University of Washington, Seattle, WA 98195-7470, USA.
Hum Mol Genet. 1998 Mar;7(3):465-9. doi: 10.1093/hmg/7.3.465.
The ATP7A gene encodes a copper-transporting ATPase. Mutations in this gene result in two clinically distinct X-linked inherited disorders: Menkes disease and occipital horn syndrome (OHS). We identified a single exon skipping in the ATP7A transcript in cells from the affected proband, affected cousins and obligate carriers in a family with OHS. Genomic sequencing identified an A-->T transversion at the +3 position in the splice donor site of intron 10 (gtaaagt-->gttaagt) in all affected individuals and the obligate female carriers. This mutation results in the constitutive skipping of exon 10 and creates an in-frame deletion of transmembrane domains 3 and 4 (78 amino acids) in the mature transcript. The exon 10-skipped transcript is present in low amounts as an alternatively spliced product in normal individuals. Immunocytochemical assay shows that these two protein products have different subcellular distributions: the major form is concentrated in the perinuclear Golgi system while the minor form (as the only form in this family with OHS) is co-localized with the endoplasmic reticulum-resident BiP protein (GRP78). These findings indicate that endoplasmic reticulum localization only of a variant ATP7A protein is insufficient to effect normal copper transport.
ATP7A基因编码一种铜转运ATP酶。该基因的突变会导致两种临床上截然不同的X连锁遗传性疾病:门克斯病和枕角综合征(OHS)。我们在一个患有OHS的家族中,在患病先证者、患病表亲和必然携带者的细胞中,发现ATP7A转录本存在单个外显子跳跃。基因组测序在所有患病个体和必然女性携带者中,在内含子10的剪接供体位点的+3位置鉴定出一个A→T颠换(gtaaagt→gttaagt)。这种突变导致外显子10的组成性跳跃,并在成熟转录本中造成跨膜结构域3和4(78个氨基酸)的框内缺失。外显子10跳跃的转录本在正常个体中作为一种可变剪接产物少量存在。免疫细胞化学分析表明,这两种蛋白质产物具有不同的亚细胞分布:主要形式集中在核周高尔基体系统,而次要形式(作为这个患有OHS的家族中的唯一形式)与内质网驻留蛋白BiP(GRP78)共定位。这些发现表明,仅变体ATP7A蛋白在内质网的定位不足以实现正常的铜转运。