Paw B H, Kaback M M, Neufeld E F
Department of Biological Chemistry, School of Medicine, University of California, Los Angeles 90024.
Proc Natl Acad Sci U S A. 1989 Apr;86(7):2413-7. doi: 10.1073/pnas.86.7.2413.
Chronic and adult-onset GM2 gangliosidoses are neurological disorders caused by marked deficiency of the A isoenzyme of beta-hexosaminidase; they occur in the Ashkenazi Jewish population, though less frequently than classic (infantile) Tay-Sachs disease. Earlier biosynthetic studies had identified a defective alpha-subunit that failed to associate with the beta-subunit. We have now found a guanosine to adenosine transition at the 3' end of exon 7, which causes substitution of serine for glycine at position 269 of the alpha-subunit [designated 269 (Gly----Ser) substitution]. An RNase protection assay was used to localize the mutation to a segment of mRNA from fibroblasts of a patient with the adult-onset disorder. That segment of mRNA (after reverse transcription) and a corresponding segment of genomic DNA were amplified by the polymerase chain reaction and sequenced by the dideoxy method. The sequence analysis, together with an assay based on the loss of a ScrFI restriction site, showed that the patient was a compound heterozygote who had inherited the 269 (Gly----Ser) mutation from his father and an allelic null mutation from his mother. The 269 (Gly----Ser) mutation, in compound heterozygosity with a presumed null allele, was also found in fetal fibroblasts with an association-defective phenotype and in cells from five patients with chronic GM2 gangliosidosis. It was not found in beta-hexosaminidase A-deficient cells obtained from patients with infantile Tay-Sachs disease nor in cells from individuals who do not have beta-hexosaminidase A deficiency. However, there must be additional mutations with similar consequences, since the 269 (Gly----Ser) substitution was not present in fibroblasts from two patients with juvenile GM2 gangliosidosis even though these had an association-defective alpha-subunit.
慢性及成人期发病的GM2神经节苷脂贮积症是一种神经疾病,由β-己糖胺酶A同工酶显著缺乏所致;该疾病在德系犹太人中出现,不过其发病率低于典型(婴儿型)泰-萨克斯病。早期的生物合成研究已鉴定出一种有缺陷的α亚基,它无法与β亚基结合。我们现在发现在外显子7的3'端有一个鸟苷到腺苷的转换,这导致α亚基第269位的甘氨酸被丝氨酸取代[称为269(甘氨酸→丝氨酸)取代]。采用核糖核酸酶保护试验将该突变定位到一名成人期发病疾病患者成纤维细胞的一段mRNA上。该段mRNA(逆转录后)和基因组DNA的相应片段通过聚合酶链反应进行扩增,并采用双脱氧法进行测序。序列分析以及基于ScrFI限制性酶切位点缺失的检测表明,该患者是一个复合杂合子,他从父亲那里继承了269(甘氨酸→丝氨酸)突变,从母亲那里继承了一个等位基因无效突变。在具有结合缺陷表型的胎儿成纤维细胞以及五名慢性GM2神经节苷脂贮积症患者的细胞中也发现了与一个假定的无效等位基因呈复合杂合状态的269(甘氨酸→丝氨酸)突变。在婴儿型泰-萨克斯病患者的β-己糖胺酶A缺陷细胞中以及在没有β-己糖胺酶A缺陷的个体细胞中均未发现该突变。然而,肯定还存在其他具有类似后果的突变,因为两名青少年GM2神经节苷脂贮积症患者的成纤维细胞中不存在269(甘氨酸→丝氨酸)取代,尽管这些细胞的α亚基存在结合缺陷。