A flow cytometry-based reporter assay identifies macrolide antibiotics as nonsense mutation read-through agents

Michal Caspi, Anastasia Firsow, Raja Rajkumar, Nir Skalka, Itay Moshkovitz, Ariel Munitz, Metsada Pasmanik-Chor, Hagar Greif, Dalia Megido, Revital Kariv, Daniel W. Rosenberg, Rina Rosin-Arbesfeld*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Abstract: A large number of human diseases are caused by nonsense mutations. These mutations result in premature protein termination and the expression of truncated, usually nonfunctional products. A promising therapeutic strategy for patients suffering from premature termination codon (PTC)-mediated disorders is to suppress the nonsense mutation and restore the expression of the affected protein. Such a suppression approach using specific antibiotics and other read-through promoting agents has been shown to suppress PTCs and restore the production of several important proteins. Here, we report the establishment of a novel, rapid, and very efficient method for screening stop-codon read-through agents. We also show that, in both mammalian cells and in a transgenic mouse model, distinct members of the macrolide antibiotic family can induce read-through of disease-causing stop codons leading to re-expression of several key proteins and to reduced disease phenotypes. Taken together, our results may help in the identification and characterization of well-needed customized pharmaceutical PTC suppression agents. Key messages: Establishment of a flow cytometry-based reporter assay to identify nonsense mutation read-through agents.Macrolide antibiotics can induce read-through of disease-causing stop codons.Macrolide-induced protein restoration can alleviate disease-like phenotypes.

Original languageEnglish
Pages (from-to)469-482
Number of pages14
JournalJournal of Molecular Medicine
Volume94
Issue number4
DOIs
StatePublished - 1 Apr 2016

Funding

FundersFunder number
Rising Tide Foundation for Clinical Cancer Research
United States-Israel Binational Science Foundation

    Keywords

    • Aminoglycoside and macrolide antibiotics
    • Genetic diseases
    • Nonsense mutations
    • Premature termination codons (PTCs)
    • Ribosomal read-through

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