Helicobacter pylori induces ERK-dependent formation of a phospho-c-Fos c-Jun activator protein-1 complex that causes apoptosis in macrophages

J Biol Chem. 2010 Jun 25;285(26):20343-57. doi: 10.1074/jbc.M110.116988. Epub 2010 Apr 21.

Abstract

Macrophages are essential components of innate immunity, and apoptosis of these cells impairs mucosal defense to microbes. Helicobacter pylori is a gastric pathogen that infects half of the world population and causes peptic ulcer disease and gastric cancer. The host inflammatory response fails to eradicate the organism. We have reported that H. pylori induces apoptosis of macrophages by generation of polyamines from ornithine decarboxylase (ODC), which is dependent on c-Myc as a transcriptional enhancer. We have now demonstrated that expression of c-Myc requires phosphorylation and nuclear translocation of ERK, which results in phosphorylation of c-Fos and formation of a specific activator protein (AP)-1 complex. Electromobility shift assay and immunoprecipitation revealed a previously unrecognized complex of phospho-c-Fos (pc-Fos) and c-Jun in the nucleus. Fluorescence resonance energy transfer demonstrated the interaction of pc-Fos and c-Jun. The capacity of this AP-1 complex to bind to putative AP-1 sequences was demonstrated by oligonucleotide pulldown and fluorescence polarization. Binding of the pc-Fos.c-Jun complex to the c-Myc promoter was demonstrated by chromatin immunoprecipitation. A dominant-negative c-Fos inhibited H. pylori-induced expression of c-Myc and ODC and apoptosis. H. pylori infection of mice induced a rapid infiltration of macrophages into the stomach. Concomitant apoptosis depleted these cells, and this was associated with formation of a pc-Fos.c-Jun complex. Treatment of mice with an inhibitor of ERK phosphorylation attenuated phosphorylation of c-Fos, expression of ODC, and apoptosis in gastric macrophages. A unique AP-1 complex in gastric macrophages contributes to the immune escape of H. pylori.

Publication types

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

MeSH terms

  • Animals
  • Anthracenes / pharmacology
  • Apoptosis*
  • Cell Line
  • Cell Nucleus / metabolism
  • Cell Survival / drug effects
  • Extracellular Signal-Regulated MAP Kinases / antagonists & inhibitors
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • Flavonoids / pharmacology
  • Fluorescence Resonance Energy Transfer
  • Helicobacter Infections / genetics
  • Helicobacter Infections / metabolism
  • Helicobacter Infections / microbiology
  • Helicobacter pylori / physiology*
  • Host-Pathogen Interactions
  • Imidazoles / pharmacology
  • Immunoblotting
  • Macromolecular Substances / metabolism*
  • Macrophages / cytology
  • Macrophages / metabolism
  • Macrophages / microbiology*
  • Mice
  • Mice, Inbred C57BL
  • Ornithine Decarboxylase / genetics
  • Ornithine Decarboxylase / metabolism
  • Phosphorylation / drug effects
  • Protein Binding
  • Proto-Oncogene Proteins c-fos / genetics
  • Proto-Oncogene Proteins c-fos / metabolism
  • Proto-Oncogene Proteins c-jun / genetics
  • Proto-Oncogene Proteins c-jun / metabolism
  • Pyridines / pharmacology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor AP-1 / genetics
  • Transcription Factor AP-1 / metabolism

Substances

  • Anthracenes
  • Flavonoids
  • Imidazoles
  • Macromolecular Substances
  • Proto-Oncogene Proteins c-fos
  • Proto-Oncogene Proteins c-jun
  • Pyridines
  • Transcription Factor AP-1
  • pyrazolanthrone
  • Extracellular Signal-Regulated MAP Kinases
  • Ornithine Decarboxylase
  • SB 203580
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one