Valerio D, Duyvesteyn M G, van Ormondt H, Meera Khan P, van der Eb A J
Nucleic Acids Res. 1984 Jan 25;12(2):1015-24. doi: 10.1093/nar/12.2.1015.
In order to determine the molecular basis of adenosine deaminase (ADA) deficiency in cells derived from patients with severe combined immunodeficiency (SCID) disease, we used a human ADA cDNA clone (1) to analyse the organization and transcription of the ADA gene in both normal and ADA-SCID cells. In five lymphoblastoid ADA-SCID cell lines we could detect no deletions or rearrangements in the ADA gene and its flanking sequences. Furthermore, synthesis and processing of ADA mRNA appeared to be normal in the ADA-SCID cells, and ADA-specific mRNA from two ADA-SCID cells could be translated in vitro into a protein with the molecular weight of normal ADA; this protein, however, could hardly be precipitated with an ADA antiserum. The results indicate that in these two ADA-SCID cell lines, the lack of ADA activity is not due to transcriptional or translational defects, but to subtle changes in the configuration of the protein affecting both its enzymatic and immunological characteristics.
为了确定重症联合免疫缺陷(SCID)病患者细胞中腺苷脱氨酶(ADA)缺乏的分子基础,我们使用人ADA cDNA克隆(1)分析正常细胞和ADA - SCID细胞中ADA基因的结构和转录情况。在五个淋巴母细胞样ADA - SCID细胞系中,我们未检测到ADA基因及其侧翼序列的缺失或重排。此外,ADA - SCID细胞中ADA mRNA的合成和加工似乎正常,来自两个ADA - SCID细胞的ADA特异性mRNA可在体外翻译成具有正常ADA分子量的蛋白质;然而,这种蛋白质几乎不能被ADA抗血清沉淀。结果表明,在这两个ADA - SCID细胞系中,ADA活性的缺乏不是由于转录或翻译缺陷,而是由于蛋白质构象的细微变化影响了其酶学和免疫学特性。