title: A flow cytometry-based reporter assay identifies macrolide antibiotics as nonsense mutation read-through agents.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel.
authors.forename: Michal
authors.initials: M
authors.lastname: Caspi
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Firsow
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel.
authors.forename: Raja
authors.initials: R
authors.lastname: Rajkumar
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel.
authors.forename: Nir
authors.initials: N
authors.lastname: Skalka
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel.
authors.forename: Itay
authors.initials: I
authors.lastname: Moshkovitz
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel.
authors.forename: Ariel
authors.initials: A
authors.lastname: Munitz
authors.affiliation: Bioinformatics Unit, G.S.W. Faculty of Life Science, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Metsada
authors.initials: M
authors.lastname: Pasmanik-Chor
authors.affiliation: Bioblast Pharma, Tel Aviv, Israel.
authors.forename: Hagar
authors.initials: H
authors.lastname: Greif
authors.affiliation: Bioblast Pharma, Tel Aviv, Israel.
authors.forename: Dalia
authors.initials: D
authors.lastname: Megido
authors.affiliation: Department of Gastroenterology, Tel Aviv Sourasky Medical Center, Tel Aviv, 64239, Israel.
authors.forename: Revital
authors.initials: R
authors.lastname: Kariv
authors.affiliation: Center for Molecular Medicine, University of Connecticut School of Medicine, Farmington, CT, USA.
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Rosenberg
authors.affiliation: Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat-Aviv, 69978, Tel Aviv, Israel. arina@post.tau.ac.il.
authors.forename: Rina
authors.initials: R
authors.lastname: Rosin-Arbesfeld
mesh_terms: D000818:Animals; D017963:Azithromycin / Q000494:pharmacology; D002460:Cell Line; D018389:Codon, Nonsense; D018388:Codon, Terminator; D004917:Erythromycin / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000187:drug effects; D017491:Genes, APC; D017930:Genes, Reporter; D006801:Humans; D007417:Intestinal Polyps / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D018942:Macrolides / Q000494:pharmacology*; D051379:Mice; D018345:Mice, Knockout; D014176:Protein Biosynthesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism
chemical_list: D018389:Codon, Nonsense; D018388:Codon, Terminator; D018942:Macrolides; D012333:RNA, Messenger; D004917:Erythromycin; D017963:Azithromycin
keywords: Aminoglycoside and macrolide antibiotics; Genetic diseases; Nonsense mutations; Premature termination codons (PTCs); Ribosomal read-through