Garbarz M, Galand C, Bibas D, Bournier O, Devaux I, Harousseau J L, Grandchamp B, Dhermy D
INSERM U409, Association Claude Bernard, Faculté de Médecine Xavier Bichat, Paris, France.
Br J Haematol. 1998 Jan;100(1):90-8. doi: 10.1046/j.1365-2141.1998.00530.x.
We studied a family with autosomal dominant hereditary spherocytosis (HS) associated with a mild spectrin deficiency. Linkage analysis using two microsatellite markers (D14S63 and D14S271) very close to the beta-spectrin gene (SPTB) showed that HS co-segregated with alleles of these microsatellite markers and the linkage between the marker and HS was statistically significant. The presence of a beta-spectrin protein polymorphism (beta-spectrin Vay; A1880V) in trans of the HS allele was not itself deleterious, but allowed the detection of decreased membrane expression of the spherocytic beta-spectrin allele in two HS-affected subjects. Direct sequencing of the coding exons of the beta-spectrin gene in one affected subject showed the presence of a G-->C transversion at the terminal nucleotide of exon 3, which did not change the leucine codon 100 (CTG-->CTC). The presence of the mutation was confirmed by restriction enzyme digestion at the DNA level in all affected SH members of the family. The G-->C mutation severely reduced the utilization of the 5' splice site and resulted in aberrant mRNA splicing with intron 3 retention.
我们研究了一个与轻度血影蛋白缺乏相关的常染色体显性遗传性球形红细胞增多症(HS)家族。使用两个与β-血影蛋白基因(SPTB)非常接近的微卫星标记(D14S63和D14S271)进行连锁分析,结果显示HS与这些微卫星标记的等位基因共分离,并且标记与HS之间的连锁具有统计学意义。在HS等位基因的反式中存在β-血影蛋白蛋白多态性(β-血影蛋白Vay;A1880V)本身并无有害影响,但使得在两名受HS影响的受试者中检测到球形红细胞β-血影蛋白等位基因的膜表达降低。对一名受影响受试者的β-血影蛋白基因编码外显子进行直接测序,结果显示在外显子3的末端核苷酸处存在G→C颠换,这并未改变亮氨酸密码子100(CTG→CTC)。通过DNA水平的限制性酶切在该家族所有受影响的SH成员中证实了该突变的存在。G→C突变严重降低了5'剪接位点的利用率,并导致异常的mRNA剪接,内含子3保留。