Jenkins P B, Abou-Alfa G K, Dhermy D, Bursaux E, Féo C, Scarpa A L, Lux S E, Garbarz M, Forget B G, Gallagher P G
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
J Clin Invest. 1996 Jan 15;97(2):373-80. doi: 10.1172/JCI118425.
We studied a French kindred with typical hereditary spherocytosis (HS). Studies of erythrocytes and erythrocyte membranes from HS individuals revealed abnormal erythrocyte membrane mechanical stability as well as 15-20% deficiency of band 3, the anion transporter. Anion transport studies of red cells from two affected individuals revealed decreased sulfate flux. Nucleotide sequence of cDNA encoding the distal third of the cytoplasmic domain and the entire transmembrane domain of band 3 obtained by RT-PCR of reticulocyte RNA of an affected family member was normal. Sequence analysis of genomic DNA from an HS individual identified a nonsense mutation of the band 3 gene, Q330X, near the end of the band 3 cytoplasmic domain. This mutation was present in genomic DNA of all HS family members and absent in DNA of unaffected family members. Using an RT-PCR-based assay, a marked quantitative decrease in accumulation of the mutant band 3 RNA was detected. Thus the codon 330 nonsense mutation is responsible for the decreased accumulation of mutant band 3 RNA and the deficiency of band 3 protein in this kindred. These results have important implications for the role of band 3 defects in the membrane pathobiology of HS as well as for the techniques used in detection of HS mutations.
我们研究了一个患有典型遗传性球形红细胞增多症（HS）的法裔家族。对HS患者的红细胞和红细胞膜进行研究，发现红细胞膜的机械稳定性异常，以及阴离子转运蛋白带3缺乏15 - 20%。对两名受影响个体的红细胞进行阴离子转运研究，发现硫酸盐通量降低。通过对一名受影响家庭成员的网织红细胞RNA进行RT-PCR获得的编码带3细胞质结构域远端三分之一和整个跨膜结构域的cDNA核苷酸序列正常。对一名HS个体的基因组DNA进行序列分析，在带3细胞质结构域末端附近发现了带3基因的无义突变Q330X。该突变存在于所有HS家族成员的基因组DNA中，而在未受影响家族成员的DNA中不存在。使用基于RT-PCR的检测方法，检测到突变带3 RNA的积累明显定量减少。因此，密码子330无义突变导致了该家族中突变带3 RNA积累减少和带3蛋白缺乏。这些结果对于带3缺陷在HS膜病理生物学中的作用以及用于检测HS突变的技术具有重要意义。
