Normal polymorphic variations and transcription of the decay accelerating factor gene in paroxysmal nocturnal hemoglobinuria cells.
Normal polymorphic variations and transcription of the decay accelerating factor gene in paroxysmal nocturnal hemoglobinuria cells.
复制标题
阵发性睡眠性血红蛋白尿细胞中腐烂加速因子基因的正常多态性变异和转录。
DOI：
10.1073/pnas.85.3.880
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发表时间：
1988
影响因子：
11.1
通讯作者：
Medof,ME
中科院分区：
文献类型：
--
作者：
Stafford,HA;Tykocinski,ML;Lublin,DM;Holers,VM;Rosse,WF;Atkinson,JP;Medof,ME
阵发性睡眠性血红蛋白尿症（PNH）是一种获得性溶血性贫血，其衰变加速因子（decay accelerating factor，C3 b）的缺乏使血细胞易于增加自体补体激活片段（C3 b）的沉积和补体介导的损伤。为探讨PNH基因缺陷的机制，我们用PNH cDNA和寡核苷酸探针对正常和PNH白细胞的DNA和mRNA进行了印迹杂交。来自正常细胞的DNA的Southern分析揭示了跨越大约等于35个DNA酶的单个基因。正常人有6种HindIII带型。在家系研究中，模式分离为三个纯合和三个杂合基因型，来自三个单倍型：A，B和C的频率分别为0.47，0.36和0.17，。寡核苷酸定位多态性HindIII位点的两个非编码区附近的外显子编码（i）蛋白质寡糖丰富的结构域和（ii）mRNA的3 '-非翻译区。8例PNH患者DAF阴性白细胞的DNA分析表明，所有研究的酶的限制性片段图谱与正常人相同。3例患者具有BC（正常= 3/32），3例患者具有AA（正常= 6/32），2例患者具有AC（正常= 8/32）HindIII基因型。三个PNH患者表现出BC基因型，两个家庭的研究表明预期的遗传模式，RNA凝胶印迹分析显示mRNA转录与正常细胞中的mRNA转录没有区别。在缺乏其他糖脂锚定蛋白的受影响PNH细胞中，没有pNH 3基因或mRNA的改变，以及pNH 3，这表明PNH细胞的病变位于糖脂锚定通路中。
In paroxysmal nocturnal hemoglobinuria (PNH), an acquired hemolytic anemia, deficiency of decay accelerating factor (DAF) renders blood cells susceptible to increased deposition of autologous complement activation fragments (C3b) and complemented-mediated injury. To investigate the mechanism of the DAF defect, DNA and mRNA from normal and PNH leukocytes were compared in blot hybridization assays by using DAF cDNA and oligonucleotide probes. Southern analyses of DNA from normal cells revealed a single gene spanning approximately equal to 35 kilobases of DNA. Six HindIII banding patterns were distinguishable among normal individuals. In family studies, the patterns segregated as three homozygous and three heterozygous genotypes deriving from three haplotypes: A, B, and C with frequencies of 0.47, 0.36, and 0.17, respectively. Oligonucleotide mapping localized the polymorphic HindIII sites to two noncoding regions in the vicinity of exons encoding (i) the protein oligosaccharide-rich domain and (ii) the mRNA 3'-untranslated region. Analyses of DNA from DAF-negative leukocytes of eight PNH patients demonstrated restriction fragment profiles identical to those of normal individuals for all enzymes studied. Three patients had the BC (normals = 3/32), three patients had the AA (normals = 6/32), and two patients had the AC (normals = 8/32) HindIII genotype. Of the three PNH patients exhibiting the BC genotype, family studies of two demonstrated the expected inheritance patterns, and RNA gel blot analyses of two showed mRNA transcripts indistinguishable from those in normal cells. The absence of DAF gene or mRNA alterations in affected PNH cells that lack other glycolipid-anchored proteins as well as DAF argues that the lesion underlying PNH cells resides in the glycolipid-anchor pathway.