title: Inhibition of Non-ATG Translational Events in Cells via Covalent Small Molecules Targeting RNA.
abstract: One major class of disease-causing RNAs is expanded repeating transcripts. These RNAs cause diseases via multiple mechanisms, including: (i) gain-of-function, in which repeating RNAs bind and sequester proteins involved in RNA biogenesis and (ii) repeat associated non-ATG (RAN) translation, in which repeating transcripts are translated into toxic proteins without use of a canonical, AUG, start codon. Herein, we develop and study chemical probes that bind and react with an expanded r(CGG) repeat (r(CGG)(exp)) present in a 5' untranslated region that causes fragile X-associated tremor/ataxia syndrome (FXTAS). Reactive compounds bind to r(CGG)(exp) in cellulo as shown with Chem-CLIP-Map, an approach to map small molecule binding sites within RNAs in cells. Compounds also potently improve FXTAS-associated pre-mRNA splicing and RAN translational defects, while not affecting translation of the downstream open reading frame. In contrast, oligonucleotides affect both RAN and canonical translation when they bind to r(CGG)(exp), which is mechanistically traced to a decrease in polysome loading. Thus, designer small molecules that react with RNA targets can be used to profile the RNAs to which they bind in cells, including identification of binding sites, and can modulate several aspects of RNA-mediated disease pathology in a manner that may be more beneficial than oligonucleotides.
authors.affiliation: †Department of Chemistry, The Scripps Research Institute, Scripps Florida, 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Wang-Yong
authors.initials: WY
authors.lastname: Yang
authors.affiliation: †Department of Chemistry, The Scripps Research Institute, Scripps Florida, 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Henry D
authors.initials: HD
authors.lastname: Wilson
authors.affiliation: †Department of Chemistry, The Scripps Research Institute, Scripps Florida, 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Sai Pradeep
authors.initials: SP
authors.lastname: Velagapudi
authors.affiliation: †Department of Chemistry, The Scripps Research Institute, Scripps Florida, 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Disney
mesh_terms: D000818:Animals; D001259:Ataxia / Q000235:genetics / Q000378:metabolism; D001710:Biotin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D005600:Fragile X Syndrome / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011132:Polyribosomes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing / Q000187:drug effects; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology*; D014202:Tremor / Q000235:genetics / Q000378:metabolism; D019680:Trinucleotide Repeat Expansion / Q000187:drug effects
chemical_list: D012322:RNA Precursors; D054852:Small Molecule Libraries; D012313:RNA; D001710:Biotin
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