无义突变会导致几种遗传性疾病,如囊性纤维化,杜氏肌营养不良症,β-地中海贫血,还有Shwachman-Diamond综合征.这些突变诱导在mRNA序列内形成提前终止密码子(PTC)。导致截短的多肽的合成。翻译连读诱导药物(TRID)介导的无意义抑制疗法是纠正这些遗传缺陷的有希望的方法。TRID会产生PTC的核糖体错误编码,称为“翻译连读”,并恢复全长和潜在功能蛋白的合成。新的恶二唑核心TRIDNV848,NV914和NV930(NV)在无义相关的体外系统中显示出翻译通读活性。在这项工作中,研究了NV分子对天然终止密码子(NTC)的可能脱靶效应。使用两种不同的体外方法来评估NV分子处理是否诱导NTC连读:(1)翻译诱导的p53分子量和功能的研究;(2)评估两种看家蛋白(Cys-C和β2M)的分子量。我们的结果表明,使用NV848,NV914或NV930的处理在两个实验系统中均未引起任何翻译改变。数据表明NV分子对PTC具有特异性作用并且对NTC具有不可检测的作用。
Nonsense mutations cause several genetic diseases such as cystic fibrosis, Duchenne muscular dystrophy, β-thalassemia, and Shwachman-Diamond syndrome. These mutations induce the formation of a premature termination codon (PTC) inside the mRNA sequence, resulting in the synthesis of truncated polypeptides.
Nonsense suppression therapy mediated by translational readthrough-inducing drugs (TRIDs) is a promising approach to correct these genetic defects. TRIDs generate a ribosome miscoding of the PTC named \"translational readthrough\" and restore the synthesis of full-length and potentially functional proteins. The new oxadiazole-core TRIDs NV848, NV914, and NV930 (NV) showed translational readthrough activity in
nonsense-related in vitro systems. In this work, the possible off-target effect of NV molecules on natural termination codons (NTCs) was investigated. Two different in vitro approaches were used to assess if the NV molecule treatment induces NTC readthrough: (1) a
study of the translational-induced p53 molecular weight and functionality; (2) the evaluation of two housekeeping proteins\' (Cys-C and β2M) molecular weights. Our results showed that the treatment with NV848, NV914, or NV930 did not induce any translation alterations in both experimental systems. The data suggested that NV molecules have a specific action for the PTCs and an undetectable effect on the NTCs.