Systematic and quantitative analysis of stop codon readthrough in Rett syndrome nonsense mutations.

IF 5.4 3区 材料科学 Q2 CHEMISTRY, PHYSICAL ACS Applied Energy Materials Pub Date : 2024-05-01 Epub Date: 2024-03-02 DOI:10.1007/s00109-024-02436-6
Dennis Lebeda, Adrian Fierenz, Lina Werfel, Rina Rosin-Arbesfeld, Julia Hofhuis, Sven Thoms
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Abstract

Rett syndrome (RTT) is a neurodevelopmental disorder resulting from genetic mutations in the methyl CpG binding protein 2 (MeCP2) gene. Specifically, around 35% of RTT patients harbor premature termination codons (PTCs) within the MeCP2 gene due to nonsense mutations. A promising therapeutic avenue for these individuals involves the use of aminoglycosides, which stimulate translational readthrough (TR) by causing stop codons to be interpreted as sense codons. However, the effectiveness of this treatment depends on several factors, including the type of stop codon and the surrounding nucleotides, collectively referred to as the stop codon context (SCC). Here, we develop a high-content reporter system to precisely measure TR efficiency at different SCCs, assess the recovery of the full-length MeCP2 protein, and evaluate its subcellular localization. We have conducted a comprehensive investigation into the intricate relationship between SCC characteristics and TR induction, examining a total of 14 pathogenic MeCP2 nonsense mutations with the aim to advance the prospects of personalized therapy for individuals with RTT. Our results demonstrate that TR induction can successfully restore full-length MeCP2 protein, albeit to varying degrees, contingent upon the SCC and the specific position of the PTC within the MeCP2 mRNA. TR induction can lead to the re-establishment of nuclear localization of MeCP2, indicating the potential restoration of protein functionality. In summary, our findings underscore the significance of SCC-specific approaches in the development of tailored therapies for RTT. By unraveling the relationship between SCC and TR therapy, we pave the way for personalized, individualized treatment strategies that hold promise for improving the lives of individuals affected by this debilitating neurodevelopmental disorder. KEY MESSAGES: The efficiency of readthrough induction at MeCP2 premature termination codons strongly depends on the stop codon context. The position of the premature termination codon on the transcript influences the readthrough inducibility. A new high-content dual reporter assay facilitates the measurement and prediction of readthrough efficiency of specific nucleotide stop contexts. Readthrough induction results in the recovery of full-length MeCP2 and its re-localization to the nucleus. MeCP2 requires only one of its annotated nuclear localization signals.

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对 Rett 综合征无义突变中终止密码子读取的系统和定量分析。
雷特综合征(RTT)是一种由甲基 CpG 结合蛋白 2(MeCP2)基因突变引起的神经发育障碍。具体来说,约 35% 的 RTT 患者的 MeCP2 基因中存在无义突变导致的过早终止密码子 (PTC)。氨基糖苷类药物可将终止密码子解释为意义密码子,从而刺激翻译通读(TR)。然而,这种治疗方法的效果取决于多个因素,包括终止密码子的类型和周围的核苷酸,统称为终止密码子上下文(SCC)。在此,我们开发了一种高含量报告系统,用于精确测量不同 SCC 下的 TR 效率、评估全长 MeCP2 蛋白的恢复及其亚细胞定位。我们对SCC特征与TR诱导之间错综复杂的关系进行了全面研究,共检测了14种致病性MeCP2无义突变,旨在推进RTT患者的个性化治疗前景。我们的研究结果表明,TR诱导可以成功地恢复全长MeCP2蛋白,尽管程度不同,这取决于SCC和PTC在MeCP2 mRNA中的特定位置。TR 诱导可导致重新建立 MeCP2 的核定位,这表明蛋白功能有可能得到恢复。总之,我们的研究结果强调了针对 SCC 的方法在开发 RTT 定制疗法中的重要性。通过揭示 SCC 与 TR 治疗之间的关系,我们为个性化的个体治疗策略铺平了道路,这些策略有望改善受这种使人衰弱的神经发育障碍影响的患者的生活。关键信息:MeCP2过早终止密码子的读通诱导效率在很大程度上取决于终止密码子的上下文。过早终止密码子在转录本上的位置会影响通读诱导率。一种新的高含量双报告检测方法有助于测量和预测特定核苷酸终止上下文的读通效率。读通诱导会导致全长 MeCP2 的恢复及其重新定位到细胞核。MeCP2 只需要一个已注释的核定位信号。
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来源期刊
ACS Applied Energy Materials
ACS Applied Energy Materials Materials Science-Materials Chemistry
CiteScore
10.30
自引率
6.20%
发文量
1368
期刊介绍: ACS Applied Energy Materials is an interdisciplinary journal publishing original research covering all aspects of materials, engineering, chemistry, physics and biology relevant to energy conversion and storage. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrate knowledge in the areas of materials, engineering, physics, bioscience, and chemistry into important energy applications.
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