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

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Název: Systematic and quantitative analysis of stop codon readthrough in Rett syndrome nonsense mutations
Autoři: Dennis Lebeda, Adrian Fierenz, Lina Werfel, Rina Rosin-Arbesfeld, Julia Hofhuis, Sven Thoms
Přispěvatelé: Lebeda, Dennis, Fierenz, Adrian, Werfel, Lina, Rosin-Arbesfeld, Rina, Hofhuis, Julia, Thoms, Sven
Zdroj: J Mol Med (Berl)
Informace o vydavateli: Springer Science and Business Media LLC, 2024.
Rok vydání: 2024
Témata: 0301 basic medicine, 0303 health sciences, 03 medical and health sciences, HEK293 Cells, Codon, Nonsense, Methyl-CpG-Binding Protein 2, Protein Biosynthesis, Rett Syndrome, Codon, Terminator, Humans, Original Article, RNA, Messenger, Methyl-CpG-Binding Protein 2/genetics [MeSH], Rare disease, Humans [MeSH], Aminoglycoside, RNA, Messenger/genetics [MeSH], Codon, Nonsense [MeSH], Protein Biosynthesis [MeSH], Translational readthrough, Rett Syndrome/metabolism [MeSH], Rett Syndrome/genetics [MeSH], Personalized medicine, Methyl-CpG-Binding Protein 2/metabolism [MeSH], HEK293 Cells [MeSH], Codon, Terminator [MeSH], Rett syndrome, RNA, Messenger/metabolism [MeSH]
Popis: 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.
Druh dokumentu: Article
Other literature type
Jazyk: English
ISSN: 1432-1440
0946-2716
DOI: 10.1007/s00109-024-02436-6
Přístupová URL adresa: https://pubmed.ncbi.nlm.nih.gov/38430393
https://pub.uni-bielefeld.de/record/2987678
https://doi.org/10.1007/s00109-024-02436-6
https://resolver.sub.uni-goettingen.de/purl?gro-2/142422
https://sfb1002.med.uni-goettingen.de/production/literature/publications/512
https://repository.publisso.de/resource/frl:6521575
Rights: CC BY
"In Copyright" Rights Statement
Přístupové číslo: edsair.doi.dedup.....ddab52863cdc5228e5669d82b5d647e8
Databáze: OpenAIRE
Popis
Abstrakt: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.
ISSN:14321440
09462716
DOI:10.1007/s00109-024-02436-6