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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Immunology

Background:

  • Sepsis involves complex transcription regulation, including activation and repression events.
  • Endotoxin tolerance, a feature of sepsis, involves epigenetic silencing in leukocytes.
  • Previous work identified an interaction between NF-kappaB transcription factor RelB and G9a in epigenetic silencing during endotoxin tolerance.

Purpose of the Study:

  • To investigate the role of dynamic nucleosome changes in sepsis phenotype.
  • To assess temporal, structural, and positional nucleosome alterations during sepsis transcription reprogramming.
  • To understand how nucleosome repositioning influences tumor necrosis factor alpha (TNFalpha) transcription in sepsis.

Main Methods:

  • Utilized a THP-1 sepsis cell model.
  • Isolated mononucleosomes via cell permeabilization and micrococcal nuclease digestion.
  • Compared TNFalpha promoter nucleosome alignment in endotoxin-responsive and -tolerant cells.

Main Results:

  • Identified dynamic nucleosome repositioning during transcription activation and silencing phases.
  • Demonstrated that nucleosome exposure of NF-kappaB sites requires NAP1 and H2A.Z for promoter opening.
  • Showed RelB-dependent endotoxin tolerance involves nucleosomes masking NF-kappaB sites, and RelB inhibition prevents this silencing.

Conclusions:

  • Nucleosome repositioning is a key mechanism controlling both induction and epigenetic silencing of TNFalpha transcription in sepsis.
  • The study highlights the plasticity of nucleosome remodeling and its dependence on RelB and ATP-dependent remodelers like BAF.
  • Findings suggest nucleosome dynamics are critical regulators of the sepsis inflammatory response.