Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA

Nat Commun. 2018 Jan 19;9(1):299. doi: 10.1038/s41467-017-02582-1.

Abstract

The ubiquitously expressed RNA-binding proteins Roquin-1 and Roquin-2 are essential for appropriate immune cell function and postnatal survival of mice. Roquin proteins repress target mRNAs by recognizing secondary structures in their 3'-UTRs and by inducing mRNA decay. However, it is unknown if other cellular proteins contribute to target control. To identify cofactors of Roquin, we used RNA interference to screen ~1500 genes involved in RNA-binding or mRNA degradation, and identified NUFIP2 as a cofactor of Roquin-induced mRNA decay. NUFIP2 binds directly and with high affinity to Roquin, which stabilizes NUFIP2 in cells. Post-transcriptional repression of human ICOS by endogenous Roquin proteins requires two neighboring non-canonical stem-loops in the ICOS 3'-UTR. This unconventional cis-element as well as another tandem loop known to confer Roquin-mediated regulation of the Ox40 3'-UTR, are bound cooperatively by Roquin and NUFIP2. NUFIP2 therefore emerges as a cofactor that contributes to mRNA target recognition by Roquin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • CD4-Positive T-Lymphocytes / cytology
  • CD4-Positive T-Lymphocytes / immunology*
  • Gene Expression Regulation
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Inducible T-Cell Co-Stimulator Protein / antagonists & inhibitors
  • Inducible T-Cell Co-Stimulator Protein / genetics*
  • Inducible T-Cell Co-Stimulator Protein / immunology
  • Inverted Repeat Sequences
  • Mice
  • Mice, Inbred C57BL
  • Nuclear Proteins / antagonists & inhibitors
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / immunology
  • Nucleic Acid Conformation
  • Primary Cell Culture
  • Protein Binding
  • RNA Stability
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • RNA-Binding Proteins / antagonists & inhibitors
  • RNA-Binding Proteins / genetics*
  • RNA-Binding Proteins / immunology
  • Receptors, OX40 / antagonists & inhibitors
  • Receptors, OX40 / genetics*
  • Receptors, OX40 / immunology
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Repressor Proteins / genetics*
  • Repressor Proteins / immunology
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Ubiquitin-Protein Ligases / genetics*
  • Ubiquitin-Protein Ligases / immunology

Substances

  • ICOS protein, human
  • Inducible T-Cell Co-Stimulator Protein
  • NUFIP2 protein, human
  • Nuclear Proteins
  • RC3H1 protein, human
  • RC3H2 protein, human
  • RNA, Small Interfering
  • RNA-Binding Proteins
  • Receptors, OX40
  • Recombinant Proteins
  • Repressor Proteins
  • TNFRSF4 protein, human
  • Ubiquitin-Protein Ligases