Binding to SMN2 pre-mRNA-protein complex elicits specificity for small molecule splicing modifiers

Nat Commun. 2017 Nov 14;8(1):1476. doi: 10.1038/s41467-017-01559-4.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biflavonoids / pharmacology
  • Cell-Free System
  • Computational Biology
  • Epoxy Compounds / pharmacology
  • Exons / genetics
  • Fibroblasts
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Ligands
  • Macrolides / pharmacology
  • Muscular Atrophy, Spinal / drug therapy*
  • Muscular Atrophy, Spinal / genetics
  • Piperazines / chemical synthesis
  • Piperazines / pharmacology*
  • Protein Binding
  • Protein Structure, Quaternary
  • Proteomics / methods
  • RNA Precursors / genetics
  • RNA Precursors / metabolism*
  • RNA Splicing / drug effects*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • RNA-Binding Proteins / metabolism*
  • Spliceosomes / drug effects
  • Spliceosomes / metabolism
  • Survival of Motor Neuron 1 Protein / genetics
  • Survival of Motor Neuron 2 Protein / genetics

Substances

  • Biflavonoids
  • Epoxy Compounds
  • Ligands
  • Macrolides
  • Piperazines
  • RNA Precursors
  • RNA, Messenger
  • RNA-Binding Proteins
  • SMN1 protein, human
  • SMN2 protein, human
  • Survival of Motor Neuron 1 Protein
  • Survival of Motor Neuron 2 Protein
  • isoginkgetin
  • pladienolide B