Kugler W, Enssle J, Hentze M W, Kulozik A E
Department Pediatrics II, University of Ulm, Germany.
Nucleic Acids Res. 1995 Feb 11;23(3):413-8. doi: 10.1093/nar/23.3.413.
Nonsense mutations of the beta-globin gene are a common cause of beta-thalassemia. It is a hallmark of these mutations not only to cause a lack of protein synthesis but also a reduction of mRNA expression. Both the pathophysiologic significance and the underlying mechanisms for this surprising phenomenon have so far remained enigmatic. We report that the reduction of the fully spliced mutant beta-globin mRNA already manifests itself within the nucleus. In contrast, the levels of mutant pre-mRNA are normal. The promoter and the 5'-untranslated region (5'-UTR) of the herpes simplex virus type 1 thymidine kinase (HSV1 Tk) gene can independently circumvent this recognition/response mechanism in cis and restore nonsense mutated beta-globin mRNA expression to normal levels. These two genetic elements can thus exert a dominant influence on the post-transcriptional control of nonsense mutated beta-globin gene expression. While wild-type mRNA levels are restored by fusion of the HSV1 Tk 5'-UTR to the nonsense mutated beta-globin reading frame, translation of a wildtype reading frame in such a hybrid is precluded. In contrast, the HSV1 Tk promoter appears to efficiently deliver the mRNA to the translational apparatus. The 5'-UTR and the promoter sequences therefore control the nuclear fate of nonsense mutated beta-globin mRNA by separable pathways. The nuclear mRNA degradation mechanisms examined here may prevent the synthesis of C-terminally truncated beta-globin chain fragments and may protect heterozygotes from clinically relevant symptoms of beta-thalassemia.
β-珠蛋白基因的无义突变是β地中海贫血的常见病因。这些突变的一个标志不仅是导致蛋白质合成缺乏,还会导致mRNA表达减少。迄今为止,这种惊人现象的病理生理意义和潜在机制仍然是个谜。我们报告,完全剪接的突变β-珠蛋白mRNA的减少在细胞核内就已显现出来。相比之下,突变前体mRNA的水平是正常的。单纯疱疹病毒1型胸苷激酶(HSV1 Tk)基因的启动子和5'-非翻译区(5'-UTR)可以独立地在顺式作用中规避这种识别/反应机制,并将无义突变的β-珠蛋白mRNA表达恢复到正常水平。因此,这两个遗传元件可以对无义突变的β-珠蛋白基因表达的转录后调控产生显性影响。虽然通过将HSV1 Tk 5'-UTR与无义突变的β-珠蛋白阅读框融合可恢复野生型mRNA水平,但在这种杂种中野生型阅读框的翻译却被阻止。相比之下,HSV1 Tk启动子似乎能有效地将mRNA传递到翻译装置。因此,5'-UTR和启动子序列通过可分离的途径控制无义突变的β-珠蛋白mRNA的核命运。这里研究的核mRNA降解机制可能会阻止C末端截短的β-珠蛋白链片段的合成,并可能保护杂合子免受β地中海贫血的临床相关症状的影响。