Chemical-Induced Read-Through at Premature Termination Codons Determined by a Rapid Dual-Fluorescence System Based on S. cerevisiae

Emiliano Altamura, Monica Borgatti, Alessia Finotti, Jessica Gasparello, Roberto Gambari, Mariangela Spinelli, Rosa Castaldo, Nicola Altamura, Georg Stoecklin
2016 PLoS ONE  
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.
more » ... 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.
doi:10.1371/journal.pone.0154260 pmid:27119736 pmcid:PMC4847774 fatcat:co4gwzq4rbfudco4ajuwp2la4i