Mutations in autosomal dominant polycystic kidney disease 2 gene: Reduced expression of PKD2 protein in lymphoblastoid cells
Mutations in autosomal dominant polycystic kidney disease 2 gene: Reduced expression of PKD2 protein in lymphoblastoid cells
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DOI：
10.1016/s0272-6386(99)70420-8
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发表时间：
1999-05-01
影响因子：
13.2
通讯作者：
del Senno, L
中科院分区：
文献类型：
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作者：
Aguiari, G;Manzati, E;del Senno, L
多囊肾病 2 (PKDP) 基因编码一种由 968 个氨基酸组成的整合膜蛋白，具有六个预测的跨膜结构域以及细胞内 NH2 和 COOH 末端，在大约 15% 的常染色体显性多囊肾病 (ADPKD) 病例中发生突变，这是一种常见的遗传性疾病，经常导致肾衰竭。为了更好地了解这种疾病的病因，我们在两个具有不同临床表型的 PKD2 连锁家族中寻找 PKD2 基因的突变。发现了常见的多态性、无义突变和移码突变。预计这两种突变都会产生 314 和 386 个氨基酸的截短蛋白质，并被捕获在蛋白质的第一个细胞外环处。聚合酶链式反应 (PCR) 和逆转录酶 (RT)-PCR 产物的限制酶分析分别表明，与疾病共分离的突变和突变等位基因在类淋巴母细胞系的信使 RNA 水平上表达。然而，在这些细胞中，蛋白质印迹分析仅显示 PKD2 正常蛋白，并且其表达水平低于没有 PKDP 突变的细胞中的表达水平。这些发现表明，在类淋巴母细胞中，突变等位基因的截短蛋白产物可能不稳定。 (C) 1999 年，国家肾脏基金会 (National Kidney Foundation, Inc.)
The polycystic kidney disease 2 (PKDP) gene, encoding a 968-amino acid integral membrane protein with six predicted membrane-spanning domains and intracellular NH2 and COOH termini, is mutated in approximately 15% of the cases of autosomal dominant polycystic kidney disease (ADPKD), a common genetic disease frequently resulting in renal failure. For a better understanding of the cause of this disorder, we searched for mutations in the PKD2 gene in two PKD2-linked families characterized by different clinical phenotypes. A common polymorphism, a nonsense mutation, and a frameshift mutation were found. Both mutations are predicted to produce truncated proteins of 314 and 386 amino acids, arrested at the first extracellular loop of the protein. Restriction enzyme analysis of polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR products, respectively, showed that mutations cosegregated with the disease and mutated alleles were expressed at the messenger RNA level in lymphoblastoid cell lines. However, in these cells, Western blot analysis showed only PKD2 normal protein, and it was expressed at a lower level than that found in cells without the PKDP mutation. These findings suggest that in lymphoblastoid cells, the truncated protein product of the mutant allele may not be stable. (C) 1999 by the National Kidney Foundation, Inc.