Recovery of TRIM25-Mediated RIG-I Ubiquitination through Suppression of NS1 by RNA Aptamers

Mol Cells. 2019 Oct 31;42(10):721-728. doi: 10.14348/molcells.2019.0157.

Abstract

Non-structural protein 1 (NS1) of influenza virus has been shown to inhibit the innate immune response by blocking the induction of interferon (IFN). In this study, we isolated two single-stranded RNA aptamers specific to NS1 with K d values of 1.62 ± 0.30 nM and 1.97 ± 0.27 nM, respectively, using a systematic evolution of ligand by exponential enrichment (SELEX) procedure. The selected aptamers were able to inhibit the interaction of NS1 with tripartite motif-containing protein 25 (TRIM25), and suppression of NS1 enabled retinoic acid inducible gene I (RIG-I) to be ubiquitinated regularly by TRIM25. Additional luciferase reporter assay and quantitative real-time PCR (RT-PCR) experiments demonstrated that suppression of NS1 by the selected aptamers induced IFN production. It is noted that viral replication was also inhibited through IFN induction in the presence of the selected aptamers. These results suggest that the isolated aptamers are strongly expected to be new therapeutic agents against influenza infection.

Keywords: aptamer; influenza virus; non-structural 1; systematic evolution of ligands by exponential enrichment; ubiquiti nation.

MeSH terms

  • Animals
  • Aptamers, Nucleotide / metabolism*
  • DEAD Box Protein 58 / metabolism*
  • HEK293 Cells
  • Humans
  • Interferons / metabolism
  • Mice
  • Protein Binding
  • RAW 264.7 Cells
  • Receptors, Immunologic
  • Transcription Factors / metabolism*
  • Tripartite Motif Proteins / metabolism*
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination*
  • Viral Nonstructural Proteins / metabolism*
  • Virus Replication

Substances

  • Aptamers, Nucleotide
  • Receptors, Immunologic
  • Transcription Factors
  • Tripartite Motif Proteins
  • Viral Nonstructural Proteins
  • Interferons
  • TRIM25 protein, human
  • Ubiquitin-Protein Ligases
  • RIGI protein, human
  • DEAD Box Protein 58