1. Binding to SMN2 pre-mRNA-protein complex elicits specificity for small molecule splicing modifiers.
- Author
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Sivaramakrishnan M, McCarthy KD, Campagne S, Huber S, Meier S, Augustin A, Heckel T, Meistermann H, Hug MN, Birrer P, Moursy A, Khawaja S, Schmucki R, Berntenis N, Giroud N, Golling S, Tzouros M, Banfai B, Duran-Pacheco G, Lamerz J, Hsiu Liu Y, Luebbers T, Ratni H, Ebeling M, Cléry A, Paushkin S, Krainer AR, Allain FH, and Metzger F
- Subjects
- Biflavonoids pharmacology, Cell-Free System, Computational Biology, Epoxy Compounds pharmacology, Exons genetics, Fibroblasts, HEK293 Cells, HeLa Cells, Humans, Ligands, Macrolides pharmacology, Muscular Atrophy, Spinal genetics, Piperazines chemical synthesis, Protein Binding, Protein Structure, Quaternary, Proteomics methods, RNA Precursors genetics, RNA, Messenger genetics, Spliceosomes drug effects, Spliceosomes metabolism, Survival of Motor Neuron 1 Protein genetics, Survival of Motor Neuron 2 Protein genetics, Muscular Atrophy, Spinal drug therapy, Piperazines pharmacology, RNA Precursors metabolism, RNA Splicing drug effects, RNA, Messenger metabolism, RNA-Binding Proteins metabolism
- Abstract
Small molecule splicing modifiers have been previously described that target the general splicing machinery and thus have low specificity for individual genes. Several potent molecules correcting the splicing deficit of the SMN2 (survival of motor neuron 2) gene have been identified and these molecules are moving towards a potential therapy for spinal muscular atrophy (SMA). Here by using a combination of RNA splicing, transcription, and protein chemistry techniques, we show that these molecules directly bind to two distinct sites of the SMN2 pre-mRNA, thereby stabilizing a yet unidentified ribonucleoprotein (RNP) complex that is critical to the specificity of these small molecules for SMN2 over other genes. In addition to the therapeutic potential of these molecules for treatment of SMA, our work has wide-ranging implications in understanding how small molecules can interact with specific quaternary RNA structures.
- Published
- 2017
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