Chemical-Induced Read-Through at Premature Termination Codons Determined by a Rapid Dual-Fluorescence System Based on S. cerevisiae |
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| Authors: | Emiliano Altamura Monica Borgatti Alessia Finotti Jessica Gasparello Roberto Gambari Mariangela Spinelli Rosa Castaldo Nicola Altamura |
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| Affiliation: | 1Chemistry Department, University of Bari, Bari, Italy;2Department of Life Sciences and Biotechnology, Biochemistry and Molecular Biology Section, University of Ferrara, Ferrara, Italy;3Institute of Biomembranes and Bioenergetics, National Researches Council, Bari, Italy;German Cancer Research Center, GERMANY |
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| Abstract: | Nonsense mutations generate in-frame stop codons in mRNA leading to a premature arrest of translation. Functional consequences of premature termination codons (PTCs) include the synthesis of truncated proteins with loss of protein function causing severe inherited or acquired diseases. A therapeutic approach has been recently developed that is based on the use of chemical agents with the ability to suppress PTCs (read-through) restoring the synthesis of a functional full-length protein. Research interest for compounds able to induce read-through requires an efficient high throughput large scale screening system. We present a rapid, sensitive and quantitative method based on a dual-fluorescence reporter expressed in the yeast Saccharomyces cerevisiae to monitor and quantitate read-through at PTCs. We have shown that our novel system works equally well in detecting read-through at all three PTCs UGA, UAG and UAA. |
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