Bessler M, Mason P J, Hillmen P, Miyata T, Yamada N, Takeda J, Luzzatto L, Kinoshita T
Department of Haematology, Hammersmith Hospital, Royal Postgraduate Medical School, London, UK.
EMBO J. 1994 Jan 1;13(1):110-7. doi: 10.1002/j.1460-2075.1994.tb06240.x.
Paroxysmal nocturnal haemoglobinuria (PNH), an acquired clonal blood disorder, is caused by the absence of glycosyl phosphatidylinositol (GPI)-anchored surface proteins due to a defect in a specific step of GPI-anchor synthesis. The cDNA of the X-linked gene, PIG-A, which encodes a protein required for this step has recently been isolated. We have carried out a molecular and functional analysis of the PIG-A gene in four cell lines deficient in GPI-linked proteins, obtained by Epstein-Barr virus (EBV) transformation of affected B-lymphocytes from PNH patients. In all four cell lines transfection with PIG-A cDNA restored normal expression of GPI-linked proteins. In three of the four cell lines the primary lesion is a frameshift mutation. In two of these there is a reduction in the amount of full-length mRNA. The fourth cell line contains a missense mutation in PIG-A. In each case the mutation was present in the affected granulocytes from peripheral blood of the patients, but not in normal sister cell lines from the same patient. These data prove that PNH is caused in most patients by a single mutation in the PIG-A gene. The nature of the mutation can vary and most likely occurs on the active X-chromosome in an early haematopoietic stem cell.
阵发性夜间血红蛋白尿(PNH)是一种获得性克隆性血液疾病,由于糖基磷脂酰肌醇(GPI)锚定表面蛋白合成特定步骤存在缺陷,导致该蛋白缺失。编码此步骤所需蛋白质的X连锁基因PIG-A的cDNA最近已被分离出来。我们对通过爱泼斯坦-巴尔病毒(EBV)转化PNH患者受影响的B淋巴细胞获得的四种缺乏GPI连接蛋白的细胞系中的PIG-A基因进行了分子和功能分析。在所有四种细胞系中,用PIG-A cDNA转染可恢复GPI连接蛋白的正常表达。在这四种细胞系中的三种中,主要病变是移码突变。其中两种细胞系中全长mRNA的量减少。第四种细胞系在PIG-A中含有错义突变。在每种情况下,突变都存在于患者外周血中受影响的粒细胞中,但不存在于同一患者的正常姐妹细胞系中。这些数据证明,大多数PNH患者是由PIG-A基因的单一突变引起的。突变的性质可能不同,最有可能发生在早期造血干细胞的活性X染色体上。