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ATF-2 regulates lipopolysaccharide-induced transcription in macrophage cells

Noriyuki Hirose1, Toshio Maekawa, Toshie Shinagawa

  • 1Laboratory of Molecular Genetics, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.

Biochemical and Biophysical Research Communications
|May 9, 2009
PubMed

Insights

The transcription factor ATF-2 regulates Toll-like receptor (TLR)-mediated gene expression in macrophages. ATF-2 is crucial for LPS-induced Socs-3 expression, involving HDAC1 interaction.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • Activating transcription factor 2 (ATF-2) is a transcription factor involved in stress responses and Toll-like receptor (TLR) signaling.
  • Macrophage activation via TLRs is critical for innate immunity and inflammatory responses.

Purpose of the Study:

  • To investigate the role of ATF-2 in TLR-mediated gene expression in RAW264.7 macrophage cells.
  • To elucidate the mechanisms by which ATF-2 regulates the expression of specific LPS-induced genes, such as Socs-3.

Main Methods:

  • RAW264.7 macrophage cells were treated with Toll-like receptor agonists (LPS, MALP-2, CpG-ODN).
  • Phosphorylation of ATF-2 and its trans-activation capacity were assessed.
  • Small interfering RNA (siRNA) targeting Atf-2 was used to evaluate its role in gene expression.
  • Chromatin immunoprecipitation (ChIP) and Western blotting were employed to study protein interactions and modifications.
  • Histone deacetylase (HDAC) activity was modulated using trichostatin A.

Main Results:

  • LPS, MALP-2, and CpG-ODN treatments enhanced ATF-2 phosphorylation at Thr-71.
  • LPS treatment increased ATF-2 trans-activation capacity.
  • Atf-2 siRNA significantly reduced LPS-induced expression of Suppressor of cytokine signaling 3 (Socs-3).
  • ATF-2 and LPS synergistically stimulated Socs-3 promoter activity, while Atf-2 siRNA suppressed it.
  • Histone deacetylase 1 (HDAC1) interacted with ATF-2 upon LPS treatment.
  • HDAC inhibition with trichostatin A suppressed LPS-induced Socs-3 expression.

Conclusions:

  • ATF-2 plays a significant role in regulating Toll-like receptor-mediated transcription in macrophages.
  • HDAC1 positively regulates LPS-induced Socs-3 transcription through interaction with ATF-2.
  • These findings highlight a novel regulatory pathway involving ATF-2 and HDAC1 in macrophage inflammatory responses.

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