Interleukin-1 homologues IL-1F7b and IL-18 contain functional mRNA instability elements within the coding region responsive to lipopolysaccharide

Biochem J. 2004 Jul 15;381(Pt 2):503-10. doi: 10.1042/BJ20040217.

Abstract

IL-1F7b, a novel homologue of the IL-1 (interleukin 1) family, was discovered by computational cloning. We demonstrated that IL-1F7b shares critical amino acid residues with IL-18 and binds to the IL-18-binding protein enhancing its ability to inhibit IL-18-induced interferon-gamma. We also showed that low levels of IL-1F7b are constitutively present intracellularly in human blood monocytes. In this study, we demonstrate that similar to IL-18, both mRNA and intracellular protein expression of IL-1F7b are up-regulated by LPS (lipopolysaccharide) in human monocytes. In stable transfectants of murine RAW264.7 macrophage cells, there was no IL-1F7b protein expression despite a highly active CMV promoter. We found that IL-1F7b-specific mRNA was rapidly degraded in transfected cells, via a 3'-UTR (untranslated region)-independent control of IL-1F7b transcript stability. After LPS stimulation, there was a rapid transient increase in IL-1F7b-specific mRNA and concomitant protein levels. Using sequence alignment, we found a conserved ten-nucleotide homology box within the open reading frame of IL-F7b, which is flanking the coding region instability elements of some selective genes. In-frame deletion of downstream exon 5 from the full-length IL-1F7b cDNA markedly increased the levels of IL-1F7b mRNA. A similar coding region element is located in IL-18. When transfected into RAW264.7 macrophages, IL-18 mRNA was also unstable unless treated with LPS. These results indicate that both IL-1F7b and IL-18 mRNA contain functional instability determinants within their coding region, which influence mRNA decay as a novel mechanism to regulate the expression of IL-1 family members.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence / genetics
  • CHO Cells / chemistry
  • CHO Cells / metabolism
  • COS Cells / chemistry
  • COS Cells / metabolism
  • Cell Line
  • Chlorocebus aethiops
  • Conserved Sequence / genetics
  • Cricetinae
  • Exons / genetics
  • Exons / physiology
  • Gene Expression Regulation / physiology
  • Humans
  • Interleukin-1 / genetics*
  • Interleukin-1 / immunology
  • Interleukin-1 / metabolism
  • Interleukin-1 / physiology
  • Interleukin-18 / genetics*
  • Interleukin-18 / immunology
  • Interleukin-18 / metabolism
  • Lipopolysaccharides / immunology*
  • Macrophages / chemistry
  • Macrophages / metabolism
  • Mice
  • Molecular Sequence Data
  • Monocytes / chemistry
  • Monocytes / metabolism
  • Monocytes / physiology
  • RNA Stability / physiology*
  • RNA, Messenger / physiology*
  • Sequence Deletion / genetics
  • Sequence Deletion / physiology
  • Sequence Homology, Nucleic Acid
  • Transfection / methods
  • Up-Regulation / physiology

Substances

  • Interleukin-1
  • Interleukin-18
  • Lipopolysaccharides
  • RNA, Messenger
  • interleukin-1 homolog 4, human