Li L, Bray P F
Department of Medicine, Johns Hopkins University Medical School, Baltimore, MD 21205.
Am J Hum Genet. 1993 Jul;53(1):140-9.
The crucial role of the human platelet fibrinogen receptor in maintaining normal hemostasis is best exemplified by the autosomal recessive bleeding disorder Glanzmann thrombasthenia (GT). The platelet fibrinogen receptor is a heterodimer composed of glycoproteins IIb (GPIIb) and IIIa (GPIIIa). Platelets from patients with GT have a quantitative or qualitative abnormality in GPIIb and GPIIIa and can neither bind fibrinogen nor aggregate. Very few genetic defects have been identified that cause this disorder. We describe a kindred with GT in which the affected individuals have a unique inversion-deletion mutation in the gene for GPIIIa. Patient platelets lacked both GPIIIa protein and mRNA. Southern blots of patient genomic DNA probed with an internal 1.0-kb GPIIIa cDNA suggested a large rearrangement of this gene but were normal when probed with small GPIIIa cDNA fragments that were outside the mutation. Cytogenetics and pulsed-field gel analysis of the GPIIIa gene were normal, making a translocation or a very large rearrangement unlikely. Additional Southern analyses suggested that the abnormality was not a small insertion. We constructed a patient genomic DNA library and isolated fragments containing the 5' and 3' breakpoints of the mutation. The nucleotide sequence from these genomic clones was determined and revealed that, relative to the normal gene, the mutant allele contained a 1-kb deletion immediately preceding a 15-kb inversion. The DNA breaks occurred in two inverted and one forward Alu sequence within the gene for GPIIIa and in the left, right, and left arms, respectively, of these sequences. There was a 5-bp repeat at the 3' terminus of the inversion.(ABSTRACT TRUNCATED AT 250 WORDS)
人类血小板纤维蛋白原受体在维持正常止血过程中的关键作用,最典型的例子是常染色体隐性出血性疾病——Glanzmann血小板无力症(GT)。血小板纤维蛋白原受体是一种异二聚体,由糖蛋白IIb(GPIIb)和IIIa(GPIIIa)组成。GT患者的血小板在GPIIb和GPIIIa上存在数量或质量异常,既不能结合纤维蛋白原也不能聚集。目前已确定的导致该疾病的基因缺陷非常少。我们描述了一个患有GT的家族,其中受影响的个体在GPIIIa基因中存在一种独特的倒位-缺失突变。患者的血小板既缺乏GPIIIa蛋白也缺乏mRNA。用1.0 kb的GPIIIa cDNA内部片段探测患者基因组DNA的Southern印迹显示该基因有大量重排,但用突变外的小GPIIIa cDNA片段探测时则正常。对GPIIIa基因进行细胞遗传学和脉冲场凝胶分析均正常,排除了易位或非常大的重排的可能性。进一步的Southern分析表明异常不是小的插入。我们构建了患者基因组DNA文库,并分离出包含突变5'和3'断点的片段。测定了这些基因组克隆的核苷酸序列,结果显示,相对于正常基因,突变等位基因在一个15 kb倒位之前紧邻一个1 kb的缺失。DNA断裂分别发生在GPIIIa基因内的两个反向和一个正向Alu序列中,以及这些序列的左臂、右臂和左臂。在倒位的3'末端有一个5 bp的重复序列。(摘要截断于250字)